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<h1 class="sectionedit1">Labjournal Surchem August</h1>
<div class="level1">
</div>
<!-- EDIT1 SECTION "Labjournal Surchem August" [1-40] -->
<h2 class="sectionedit2">Experiment 63e: Anti Mouse/Rabbit on PDITC</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT2 SECTION "Experiment 63e: Anti Mouse/Rabbit on PDITC" [41-217] -->
<h3 class="sectionedit3">2015.08.29</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 3 PDITC slides were prepared</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
<li class="level1"><div class="li"> desiccator: 15 min</div>
</li>
<li class="level1"><div class="li"> stored at 4 °C</div>
</li>
</ul>
</div>
<!-- EDIT3 SECTION "2015.08.29" [218-435] -->
<h3 class="sectionedit4">2015.08.31</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<p>
spotting pattern:
<img alt="" class="media" src="https://static.igem.org/mediawiki/2015/8/8a/Freiburg_labjournal-surchem-spotting_pattern_3.png" width="300"/><br/>

slides: 424, 012
</p>
<div class="table sectionedit5"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1 </td><td class="col1"> GFP </td><td class="col2"> 0.5 mg/ml </td>
</tr>
<tr class="row2">
<td class="col0"> 2 </td><td class="col1"> anti HCV (rabbit) </td><td class="col2"> 0.5 mg/ml </td>
</tr>
<tr class="row3">
<td class="col0"> 3 </td><td class="col1"> anti Tetanus (mouse) </td><td class="col2"> 0.5 mg/ml </td>
</tr>
</table></div>
<!-- EDIT5 TABLE [581-717] -->
<p>
slide: 254
</p>
<div class="table sectionedit6"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1 </td><td class="col1"> GFP </td><td class="col2"> 1 mg/ml </td>
</tr>
<tr class="row2">
<td class="col0"> 2 </td><td class="col1"> anti HCV (rabbit) </td><td class="col2"> 1 mg/ml </td>
</tr>
<tr class="row3">
<td class="col0"> 3 </td><td class="col1"> anti Tetanus (mouse) </td><td class="col2"> 1 mg/ml </td>
</tr>
</table></div>
<!-- EDIT6 TABLE [730-860] -->
<p>
Flush protocol:
</p>
<div class="table sectionedit7"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">anti-GFP</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">anti- rabbit </td><td class="col1">6</td><td class="col2">30</td><td class="col3" colspan="2">600</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">anti- mouse </td><td class="col1">8</td><td class="col2">30</td><td class="col3" colspan="2">600</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT7 TABLE [879-1157] -->
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/70/Freiburg_labjournal-surchem-exp_63e_slide_012.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63e_slide_012.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 012: Quotion picture</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/b/b3/Freiburg_labjournal-surchem-exp_63e_slide_012_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63e_slide_012_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 012: binding curves</div></div></div><hr/>
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/0/0d/Freiburg_labjournal-surchem-exp_63e_slide_424.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63e_slide_424.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 424: Quotion picture</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/1/1f/Freiburg_labjournal-surchem-exp_63e_slide_424_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63e_slide_424_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 424: binding curves</div></div></div><hr/>
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/fa/Freiburg_labjournal-surchem-exp_63e_slide_257.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63e_slide_257.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 254: Quotion picture</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/0/06/Freiburg_labjournal-surchem-exp_63e_slide_257_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63e_slide_257_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 254: binding curves</div></div></div>
<p>
→ the detection of GFP-, rabbit-, and mouse antibodies worked on all three slides 
</p>
<p>
→ the increase in protein concentration of the spotted antigens on slide 254 does not lead to a increase of the detection signal → surface is with a protein concentration of 500 µg/ml already saturated
</p>
</div>
<!-- EDIT4 SECTION "2015.08.31" [436-2018] -->
<h2 class="sectionedit8">Experiment 63d: Stockholm II</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT8 SECTION "Experiment 63d: Stockholm II" [2019-2180] -->
<h3 class="sectionedit9">2015.08.29</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 1 PDITC slides were prepared</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
<li class="level1"><div class="li"> desiccator: 15 min</div>
</li>
<li class="level1"><div class="li"> stored at 4 °C</div>
</li>
</ul>
</div>
<!-- EDIT9 SECTION "2015.08.29" [2181-2398] -->
<h3 class="sectionedit10">2015.08.30</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<p>
spotting pattern:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<p>
slide-#:315
</p>
<div class="table sectionedit11"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1-3 </td><td class="col1">rhErbB2/Fc</td><td class="col2">100 µg/ml </td>
</tr>
<tr class="row2">
<td class="col0"> 4 </td><td class="col1"> GFP (desalt)</td><td class="col2"> 1 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0"> 5</td><td class="col1"> Tetanus antigene</td><td class="col2"> 100 µg/ml</td>
</tr>
</table></div>
<!-- EDIT11 TABLE [2542-2668] -->
</div>
<!-- EDIT10 SECTION "2015.08.30" [2399-2669] -->
<h3 class="sectionedit12">2015.08.31</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<p>
Flush protocol:
</p>
<div class="table sectionedit13"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">anti-GFP</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">affibody </td><td class="col1">6</td><td class="col2">20</td><td class="col3">900</td><td class="col4">diluted 1:3</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">affibody</td><td class="col1">8</td><td class="col2">20</td><td class="col3">900</td><td class="col4">undiluted</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row10">
<td class="col0">Anti His</td><td class="col1">10</td><td class="col2">30</td><td class="col3">600</td><td class="col4">undiluted</td>
</tr>
<tr class="row11">
<td class="col0">Buffer</td><td class="col1">11</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT13 TABLE [2735-3078] -->
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/b/be/Freiburg_labjournal-surchem-exp_63d_slide315_overview.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63d_slide315_overview.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 315: Quotion picture</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/f1/Freiburg_labjournal-surchem-exp_63d_slide315.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63d_slide315.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 315: binding curves</div></div></div>
<p>
→ Binding of the anti-his antibody to the antigen (rhErbB2/Fc) was detected→ the (His tagged) antigen was immobilized on the surface
</p>
<p>
→ no binding of the affibody to its antigen couold be detected → either the affibody does not bind strong enough or the affibody is too small (~8 kDa) to generate a sufficient iRIf signal
</p>
</div>
<!-- EDIT12 SECTION "2015.08.31" [2670-3601] -->
<h2 class="sectionedit14">Experiment 63c: Tetanus on PDITC</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT14 SECTION "Experiment 63c: Tetanus on PDITC" [3602-3770] -->
<h3 class="sectionedit15">2015.08.29</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 2 PDITC slides were prepared</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
<li class="level1"><div class="li"> desiccator: 15 min</div>
</li>
<li class="level1"><div class="li"> stored at 4 °C</div>
</li>
</ul>
</div>
<!-- EDIT15 SECTION "2015.08.29" [3771-3988] -->
<h3 class="sectionedit16">2015.08.30</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<p>
Spotting pattern:<img alt="" class="media" src="https://static.igem.org/mediawiki/2015/8/8a/Freiburg_labjournal-surchem-spotting_pattern_3.png" width="300"/><br/>

Slides: 303,466
</p>
<div class="table sectionedit17"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1 </td><td class="col1"> pos. control (GFP) </td><td class="col2"> 0.5 mg/ml </td>
</tr>
<tr class="row2">
<td class="col0"> 2 </td><td class="col1"> neg. con (Salmonella) </td><td class="col2"> (1.24 mg/ml) desalt 1:3 diluted </td>
</tr>
<tr class="row3">
<td class="col0"> 3 </td><td class="col1"> Tetanus </td><td class="col2"> (0.46 mg/ml) desalt undiluted </td>
</tr>
</table></div>
<!-- EDIT17 TABLE [4132-4317] -->
</div>
<!-- EDIT16 SECTION "2015.08.30" [3989-4318] -->
<h3 class="sectionedit18">2015.08.31</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<p>
Flush protocol:
</p>
<div class="table sectionedit19"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">anti-GFP</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">Serum (-/+) Sig002</td><td class="col1">6</td><td class="col2">30</td><td class="col3">900</td><td class="col4">diluted in 5 mg/ml BSA in PBS</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT19 TABLE [4385-4651] -->
<p>
Slide 303: flushed with Serum (-) diluted 1:20 in BSA<br/>

Slide 466: flushed with Serum (-) diluted 1:30 in BSA
</p>
</div>
<h4>Results</h4>
<div class="level4">
<p>
serum (-) 1:20 dilution:
</p>
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/a/ab/Freiburg_labjournal-surchem-exp_63c_slide303.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63c_slide303.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 303: Quotion picture</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/7b/Freiburg_labjournal-surchem-exp_63c-slide303_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63c-slide303_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 303: binding curves</div></div></div>
<p>
→ strong binding to the salmonella antigen was detected → SIG 002 could also experienced a previous salmonella infection. Signal overall was quite weak.  
</p>
<hr/>
<p>
serum (-) 1:30 dilution:
</p>
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/60/Freiburg_labjournal-surchem-exp_63c_slide466.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63c_slide466.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 466: Quotion picture</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/e/e9/Freiburg_labjournal-surchem-exp_63c_slide466-binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63c_slide466-binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 466: binding curves</div></div></div>
<p>
→the 1:30 diluted serum(-) shows little unspecific binding → weak binding to tetanus antigen could be caused by low anti tetanus antibody titer from previous vaccination
</p>
<p>
→overall weak signal, reuslts rather inconsistent→ use new stock of blood serum for previous experiments
</p>
</div>
<!-- EDIT18 SECTION "2015.08.31" [4319-5623] -->
<h2 class="sectionedit20">Experiment 63b: Ni-NTA iRIf slides</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT20 SECTION "Experiment 63b: Ni-NTA iRIf slides" [5624-5793] -->
<h3 class="sectionedit21">2015.08.29</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 4 PDITC slides were prepared</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
<li class="level1"><div class="li"> slides were sandwiched with NTA-solution and incubated o/n at 4 °C</div>
</li>
</ul>
</div>
<!-- EDIT21 SECTION "2015.08.29" [5794-6040] -->
<h3 class="sectionedit22">2015.08.30</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocked slides with APTES blocking solution for 1 h</div>
</li>
<li class="level1"><div class="li"> washed slides 2x with PBS for 10 min</div>
</li>
<li class="level1"><div class="li"> incubated slides for 1 h in 1 % NiSO<sub>4</sub>-solution</div>
</li>
<li class="level1"><div class="li"> washed slides with PBS/2x diluted PBS</div>
</li>
<li class="level1"><div class="li"> stored slides at 4 °C under N<sub>2</sub>-atmosphere</div>
</li>
</ul>
</div>
<!-- EDIT22 SECTION "2015.08.30" [6041-6348] -->
<h2 class="sectionedit23">Experiment 63a: Salmonella antigene on PDITC in iRIf</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT23 SECTION "Experiment 63a: Salmonella antigene on PDITC in iRIf" [6349-6536] -->
<h3 class="sectionedit24">2015.08.29</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 4 PDITC slides were prepared</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
</ul>
<p>
spotting pattern:<img alt="" class="media" src="https://static.igem.org/mediawiki/2015/8/8a/Freiburg_labjournal-surchem-spotting_pattern_3.png" width="300"/>
</p>
<p>
slide-#: 208, 216, 423, 500 
</p>
<div class="table sectionedit25"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1 </td><td class="col1"> pos. control (GFP) </td><td class="col2"> 1 mg/ml </td>
</tr>
<tr class="row2">
<td class="col0"> 2 </td><td class="col1"> neg. control (mCherry) </td><td class="col2"> 1mg/ml </td>
</tr>
<tr class="row3">
<td class="col0"> 3 </td><td class="col1"> Salmonella Antigen (15) </td><td class="col2"> undiluted </td>
</tr>
</table></div>
<!-- EDIT25 TABLE [6817-6970] -->
</div>
<!-- EDIT24 SECTION "2015.08.29" [6537-6975] -->
<h3 class="sectionedit26">2015.08.30</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA in PBS for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked with 10 mg/mL BSA in PBS for 30 min</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS 3x 10 min</div>
</li>
<li class="level1"><div class="li"> slides were washed with Aqua dest and dried</div>
</li>
</ul>
<p>
Flush protocol:
</p>
<div class="table sectionedit27"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">anti-GFP</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">anti-Salmonella (1301, desalt)</td><td class="col1">6</td><td class="col2">30</td><td class="col3">900</td><td class="col4">1:2 Dilution in 5 mg/ml BSA in PBS</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT27 TABLE [7254-7537] -->
<p>
208: with PBS 423: with TBS 216: with TBS 500: with TBS (changed flush protocol: 1 ug/ml a-GFP; and used anti-Salmonella lysate 1:2 diluted in 5 mg/ml BSA in TBS) 
</p>
</div>
<h4>Results</h4>
<div class="level4">
<p>
PBS:
</p>
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/e/e2/Freiburg_labjournal-surchem-exp_63a_slide208.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63a_slide208.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 208: Quotion picture</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/5/5f/Freiburg_labjournal-surchem-exp_63a_slide208_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63a_slide208_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 208: binding curves</div></div></div>
<p>
→ only very weak Salmonella antigen/antibody binding detectable, probably due to old salmonella antigen and antibody stocks
</p>
<hr/>
<p>
TBS:
</p>
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/a/a6/Freiburg_labjournal-surchem-exp_63a_slide216.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63a_slide216.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 216: Quotion picture</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/f8/Freiburg_labjournal-surchem-exp_63a_slide216_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63a_slide216_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 216: binding curves</div></div></div><div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/c/cd/Freiburg_labjournal-surchem-exp_63a_slide423.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63a_slide423.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 423: Quotion picture</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/28/Freiburg_labjournal-surchem-exp_63a_slide423_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63a_slide423_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 423: binding curves</div></div></div>
<p>
→ only very weak or no Salmonella antigen/antibody binding detectable, probably due to old salmonella antigen and antibody stocks
</p>
<hr/>
<p>
Salmonella Lysate:
</p>
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/a/a0/Freiburg_labjournal-surchem-exp_63a_slide500.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63a_slide500.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 500: Quotion picture</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/42/Freiburg_labjournal-surchem-exp_63a_slide500_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_63a_slide500_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 500: binding curves</div></div></div>
<p>
→ much unspecific binding detected, probably due to binding of E. coli proteins of the salmonella antibody lysate to other E. coli proteins still present in the purified mCherry and salmonella antigen (both were expressed in E. coli)
</p>
</div>
<!-- EDIT26 SECTION "2015.08.30" [6976-8963] -->
<h2 class="sectionedit28">Experiment 62c: PDITC/Ni-NTA slides for iRIf</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT28 SECTION "Experiment 62c: PDITC/Ni-NTA slides for iRIf" [8964-9143] -->
<h3 class="sectionedit29">2015.08.26</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 4 PDITC slides were prepared</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 1 h</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 3 h</div>
</li>
<li class="level1"><div class="li"> slides were stored under N<sub>2</sub>-atmosphere at 4 °C</div>
</li>
</ul>
</div>
<!-- EDIT29 SECTION "2015.08.26" [9144-9378] -->
<h2 class="sectionedit30">Experiment 62b:Tetanus antigen on PDITC slides for iRIf</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT30 SECTION "Experiment 62b:Tetanus antigen on PDITC slides for iRIf" [9379-9569] -->
<h3 class="sectionedit31">2015.08.26</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 4 PDITC slides were prepared</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 1 h</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 3 h</div>
</li>
<li class="level1"><div class="li"> slides incubated o/n at 4 °C</div>
</li>
</ul>
</div>
<!-- EDIT31 SECTION "2015.08.26" [9570-9776] -->
<h3 class="sectionedit32">2015.08.29</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spotted Tetanus antigen  </div>
</li>
</ul>
<p>
spotting pattern:<img alt="" class="media" src="https://static.igem.org/mediawiki/2015/8/8a/Freiburg_labjournal-surchem-spotting_pattern_3.png" width="300"/>
</p>
<p>
slide-#: 287, 024, 426, 443
</p>
<div class="table sectionedit33"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1 </td><td class="col1"> pos. control (GFP) </td><td class="col2"> 1 mg/ml </td>
</tr>
<tr class="row2">
<td class="col0"> 2 </td><td class="col1"> neg. control (Salmonella antigen) </td><td class="col2"> desalt 1:3 </td>
</tr>
<tr class="row3">
<td class="col0"> 3 </td><td class="col1"> Tetanus antigen </td><td class="col2"> undiluted </td>
</tr>
</table></div>
<!-- EDIT33 TABLE [9961-10121] -->
</div>
<!-- EDIT32 SECTION "2015.08.29" [9777-10122] -->
<h3 class="sectionedit34">2015.08.30</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA in PBS for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked with 10 mg/mL BSA in PBS for 30 min</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS and 2x with 1/10 diluted PBS for 10 min</div>
</li>
<li class="level1"><div class="li"> slides were dried and measured in iRIf</div>
</li>
</ul>
<p>
Flush protocol:
</p>
<div class="table sectionedit35"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">anti-GFP</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">Serum (-/+) Sig002</td><td class="col1">6</td><td class="col2">30</td><td class="col3">900</td><td class="col4">1:10 diluted in 5 mg/ml BSA in PBS</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT35 TABLE [10426-10697] -->
<p>
Slides: 287, 024 (with Serum (-)) #287: Serum (-) was diluted only 1:10; changed to 1:30 because Serum(-) quite high Signal at negative control and Tetanus spot Flush protocol
Slides: 426,443 (with Serum (+)) Flush protocol
</p>
</div>
<h4>Results</h4>
<div class="level4">
<p>
Serum (-):
</p>
<div class="thumb2 trien" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/9/94/Freiburg_labjournal-surchem-exp_62b_slide024.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_62b_slide024.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 024: Quotion picture</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/0/00/Freiburg_labjournal-surchem-exp_62b_slide024_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_62b_slide024_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 024: binding curves</div></div></div><div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/66/Freiburg_labjournal-surchem-exp_62b_slide287.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_62b_slide287.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 287: ROI selection</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/1/1f/Freiburg_labjournal-surchem-exp_62b_slide287_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_62b_slide287_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 287: binding curves</div></div></div>
<p>
→ slide 024 shows low unspecific binding due to 1:30 dilution of the serum(-), slide 287 shows high unspecific binding due to the 1:10 dilution of the serum(-)
</p>
<p>
→overall signal much weaker than before, probably due to repeated freezing of the blood serum
</p>
<hr/>
<p>
Serum (+):
</p>
<div class="thumb2 trien" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/5/57/Freiburg_labjournal-surchem-exp_62b_slide426.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_62b_slide426.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 426: ROI selection</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/4a/Freiburg_labjournal-surchem-exp_62b_slide426_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_62b_slide426_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 426: binding curves</div></div></div><div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/c/c1/Freiburg_labjournal-surchem-exp_62b_slide433.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_62b_slide433.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 433: ROI selection</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/4d/Freiburg_labjournal-surchem-exp_62b_slide433_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_62b_slide433_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 433: binding curves</div></div></div>
<p>
→slide 426 shows relativly weak unspecific binding and moderate binding of proteins to the Tetanus antigen, slide 433 has more unspecific binding 
</p>
<p>
→overall signal much weaker than before, probably due to repeated freezing of the blood serum
</p>
</div>
<!-- EDIT34 SECTION "2015.08.30" [10123-12172] -->
<h2 class="sectionedit36">Experiment 62a: PDITC/Ni-NTA slides for iRIf - Stockholm collaboration</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT36 SECTION "Experiment 62a: PDITC/Ni-NTA slides for iRIf - Stockholm collaboration" [12173-12378] -->
<h3 class="sectionedit37">2015.08.26</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 2 PDITC slides were prepared</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 1 h</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 3 h</div>
</li>
<li class="level1"><div class="li"> slides were sancwiched with NTA-solution and incubated o/n at 4 °C</div>
</li>
</ul>
</div>
<!-- EDIT37 SECTION "2015.08.26" [12379-12622] -->
<h3 class="sectionedit38">2015.08.27</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocked slides with APTES blocking solution for 1 h</div>
</li>
<li class="level1"><div class="li"> washed slides 2x with PBS for 10 min</div>
</li>
<li class="level1"><div class="li"> incubated slides for 1 h in 1 % NiSO<sub>4</sub>-solution</div>
</li>
<li class="level1"><div class="li"> washed slides with PBS/2x diluted PBS</div>
</li>
<li class="level1"><div class="li"> stored slides at 4 °C under N<sub>2</sub>-atmosphere</div>
</li>
</ul>
</div>
<!-- EDIT38 SECTION "2015.08.27" [12623-12931] -->
<h3 class="sectionedit39">2015.08.29</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spotted 3 µL of affibody (from Stockholm) and controls (GFP, bBSA)</div>
</li>
</ul>
<p>
spotting pattern:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<p>
slide-#: <strong>500</strong>
</p>
<div class="table sectionedit40"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1-3 </td><td class="col1"> 200 mL culture (blue heart) Lysate</td><td class="col2"> pure </td>
</tr>
<tr class="row2">
<td class="col0"> 4-6 </td><td class="col1"> 200 mL culture (blue heart) Lysate</td><td class="col2"> 1:2 diluted </td>
</tr>
<tr class="row3">
<td class="col0"> 7-9</td><td class="col1"> 10 mL culture (red heart) Lysate</td><td class="col2"> pure </td>
</tr>
<tr class="row4">
<td class="col0"> 10 </td><td class="col1 leftalign"> His-GFP desalt.  </td><td class="col2"> ~1 mg/mL </td>
</tr>
<tr class="row5">
<td class="col0"> 11 </td><td class="col1"> bBSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
</table></div>
<!-- EDIT40 TABLE [13153-13404] -->
</div>
<h4>Results</h4>
<div class="level4">
<p>
→ because of the His tag of the antigen it binds to the Ni-NTA surface→no specific antigen/affibody binding was detectable<br/>

→ retry the experiment with immobilized antigen on a PDITC surface, flush the affibody through the flow camber 
</p>
</div>
<!-- EDIT39 SECTION "2015.08.29" [12932-13659] -->
<h2 class="sectionedit41">Experiment 61: DNA on PDMS 6.0</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/PDMS_PDITC" title="protocols:pdms_pditc">PDMS-PDITC</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Cellfex_PDMS" title="protocols:cellfree_expression_on_pdms">DNA + Cellfree Expression on PDMS</a></div>
</li>
</ul>
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 3 PDMS slides for spotting with DNA and cell-free expression</div>
</li>
</ul>
</div>
<!-- EDIT41 SECTION "Experiment 61: DNA on PDMS 6.0" [13660-13984] -->
<h3 class="sectionedit42">2015.08.25</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> used 2 flow cells made on 18.08.2015: one with same pattern as flow cells used in iRIf (conic, one hole) and one with three holes and cavities, +1 made on 16.08.15 with Standard flow cell form and small borders</div>
</li>
<li class="level1"><div class="li"> washed PDMS slides with Ethanol and H<sub>2</sub>O</div>
</li>
<li class="level1"><div class="li"> activated slides in plasma generator with 30 L/h for 1 min</div>
</li>
<li class="level1"><div class="li"> incubated slides in APTES o/n at room temperature</div>
</li>
</ul>
</div>
<!-- EDIT42 SECTION "2015.08.25" [13985-14419] -->
<h3 class="sectionedit43">2015.08.26</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> Flow cells were washed individually with ethanol</div>
</li>
<li class="level1"><div class="li"> Washing step of 5 min in ethanol in slideholder</div>
</li>
<li class="level1"><div class="li"> Flow cells were dried with wafergun</div>
</li>
<li class="level1"><div class="li"> heated 90 min in oven at 70°C</div>
</li>
<li class="level1"><div class="li"> cooled down with wafergun</div>
</li>
<li class="level1"><div class="li"> Flow cells were incubated in PDITC for 2h</div>
</li>
<li class="level1"><div class="li"> filled slideholder with EtOH and discarded it immediately</div>
</li>
<li class="level1"><div class="li"> washed 2x 5 min with EtOH in slideholder</div>
</li>
<li class="level1"><div class="li"> filled slideholder again with EtOH and discarded it immediately</div>
</li>
<li class="level1"><div class="li"> dried with wafergun</div>
</li>
<li class="level1"><div class="li"> stored at 4°C in nitrogen atmosphere</div>
</li>
</ul>
</div>
<!-- EDIT43 SECTION "2015.08.26" [14420-14964] -->
<h3 class="sectionedit44">2015.08.27</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots of DNA were put on flow cells according to the following pattern</div>
</li>
<li class="level1"><div class="li"> incubation overnight at room temperature in humid atmosphere</div>
</li>
</ul>
<p>
spotting pattern (slide 61a):
</p>
<div class="table sectionedit45"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> sample </th><th class="col2"> concentration </th><th class="col3"> spot size </th>
</tr>
<tr class="row1">
<td class="col0">1</td><td class="col1">HA-GFP-Halo-His-His (Tobi)</td><td class="col2">25 ng/µl</td><td class="col3">3 µl</td>
</tr>
<tr class="row2">
<td class="col0">2</td><td class="col1">HA-GFP-Halo-His-His (Tobi)</td><td class="col2">25 ng/µl</td><td class="col3">5 µl</td>
</tr>
<tr class="row3">
<td class="col0">3</td><td class="col1">Cy3-HA-GFP-His-His</td><td class="col2">33 ng/µl</td><td class="col3">4 µl</td>
</tr>
<tr class="row4">
<td class="col0">4</td><td class="col1">Cy3-HA-GFP-His-His</td><td class="col2">66.1 ng/µl</td><td class="col3">4 µl</td>
</tr>
</table></div>
<!-- EDIT45 TABLE [15184-15400] -->
<p>
spotting pattern (slide 61b):
</p>
<div class="table sectionedit46"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> sample </th><th class="col2"> concentration </th><th class="col3"> spot size </th>
</tr>
<tr class="row1">
<td class="col0">1</td><td class="col1">Cy3-HA-GFP-His-His</td><td class="col2">66.1 ng/µl</td><td class="col3">5 µl</td>
</tr>
<tr class="row2">
<td class="col0">2</td><td class="col1">HA-GFP-Halo-His-His (Tobi)</td><td class="col2">99 ng/µl</td><td class="col3">3 µl</td>
</tr>
<tr class="row3">
<td class="col0">3</td><td class="col1">Cy3-HA-GFP-His-His</td><td class="col2">66.1 ng/µl</td><td class="col3">10 µl</td>
</tr>
</table></div>
<!-- EDIT46 TABLE [15433-15605] -->
<p>
spotting pattern (slide 61c):
</p>
<div class="table sectionedit47"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> sample </th><th class="col2"> concentration </th><th class="col3"> spot size </th>
</tr>
<tr class="row1">
<td class="col0">1</td><td class="col1">Cy3-HA-GFP-His-His</td><td class="col2">66.1 ng/µl</td><td class="col3">3 µl</td>
</tr>
<tr class="row2">
<td class="col0">2</td><td class="col1">Cy3-HA-GFP-His-His</td><td class="col2">33 ng/µl</td><td class="col3">4 µl</td>
</tr>
<tr class="row3">
<td class="col0">3</td><td class="col1">HA-GFP-Halo-His-His (Tobi)</td><td class="col2">25 ng/µl</td><td class="col3">3 µl</td>
</tr>
<tr class="row4">
<td class="col0">4</td><td class="col1">HA-GFP-Halo-His-His (Tobi)</td><td class="col2">25 ng/µl</td><td class="col3">5 µl</td>
</tr>
</table></div>
<!-- EDIT47 TABLE [15638-15854] --><ul>
<li class="level1"><div class="li"> checked for <strong>Cy3-intensity</strong> by spotting a 1.5 µl spot of Cy3-labeled GFP (33 ng/µl) on old PDMS flow cell from 18.08.2015</div>
</li>
<li class="level1"><div class="li"> spot was let dry in at room temperature until its borders were still visible</div>
</li>
<li class="level1"><div class="li"> checked for fluorescence in Mircoarray Scanner</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> Cy3 spot could be detected</div>
</li>
</ul>
</div>
<!-- EDIT44 SECTION "2015.08.27" [14965-16166] -->
<h3 class="sectionedit48">2015.08.28</h3>
<div class="level3">
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
slide 254: Retikulozyte, slide 012: Roth, slide 423: Koko
</p>
<div class="table sectionedit49"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">450</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">450</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">450</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">anti-GFP</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">strep-cy5</td><td class="col1">6</td><td class="col2">60</td><td class="col3">300</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT49 TABLE [16266-16501] -->
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/2a/Freiburg_labjournal-platereader_20150828_12.jpg" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:platereader_20150828_12.jpg" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 12 (expressed with AG Roth mix): Platereader cy5 signal after iRIf</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/c/c7/Freiburg_labjournal-roi_selection_20150828_12.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:roi_selection_20150828_12.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 12 (expressed with AG Roth mix): ROI selection </div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/1/11/Freiburg_labjournal-binding_curve_20150828_12.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:binding_curve_20150828_12.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 12 (expressed with AG Roth mix): Binding curves</div></div></div><hr/>
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/b/bf/Freiburg_labjournal-platereader_20150828_254.jpg" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:platereader_20150828_254.jpg" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 254 (expressed with retikulozyte mix): Platereader cy5 signal after iRIf</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/f9/Freiburg_labjournal-roi_selection_20150828_254.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:roi_selection_20150828_254.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 254 (expressed with retikulozyte mix): ROI selection</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/8/83/Freiburg_labjournal-binding_curve_20150828_254.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:binding_curve_20150828_254.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 254 (expressed with retikulozyte mix): Binding curves</div></div></div><hr/>
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/78/Freiburg_labjournal-platereader_20150828_423.jpg" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:platereader_20150828_423.jpg" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 423 (expressed with Koko mix): Platereader cy5 signal after iRIf</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/25/Freiburg_labjournal-roi_selection_20150828_423.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:roi_selection_20150828_423.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 423 (expressed with Koko mix): ROI selection</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/fb/Freiburg_labjournal-binding_curve_20150828_423.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:binding_curve_20150828_423.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>423 (expressed with Koko mix): Binding curves</div></div></div>
</div>
<!-- EDIT48 SECTION "2015.08.28" [16167-17610] -->
<h2 class="sectionedit50">Experiment 60: DNA on PDMS 5.0</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/PDMS_PDITC" title="protocols:pdms_pditc">PDMS-PDITC</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Cellfex_PDMS" title="protocols:cellfree_expression_on_pdms">DNA + Cellfree Expression on PDMS</a></div>
</li>
</ul>
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 PDMS slides for spotting with DNA</div>
</li>
</ul>
</div>
<!-- EDIT50 SECTION "Experiment 60: DNA on PDMS 5.0" [17611-17910] -->
<h3 class="sectionedit51">2015.08.23</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> used 2 flow cells made on 18.08.2015: one with same pattern as flow cells used in iRIf (conic, one hole) and one with three holes and cavities</div>
</li>
<li class="level1"><div class="li"> washed PDMS slides with Ethanol and H<sub>2</sub>O</div>
</li>
<li class="level1"><div class="li"> activated slides in plasma generator with 30 L/h for 1 min</div>
</li>
<li class="level1"><div class="li"> incubated slides in APTES o/n at room temperature</div>
</li>
</ul>
</div>
<!-- EDIT51 SECTION "2015.08.23" [17911-18277] -->
<h3 class="sectionedit52">2015.08.24</h3>
<div class="level3">
<ul>
<li class="level1"><div class="li"> each slide was washed with ethanol</div>
</li>
<li class="level1"><div class="li"> a 5 min washing step in Ethanol followed</div>
</li>
<li class="level1"><div class="li"> after drying with wafergun slides were baked in oven (old one) for 120 min at 70°C</div>
</li>
<li class="level1"><div class="li"> slides were cooled down with wafergun</div>
</li>
<li class="level1"><div class="li"> incubation in PDITC for 3h30 at RT</div>
</li>
<li class="level1"><div class="li"> filled slideholder with EtOH and discarded it immediately</div>
</li>
<li class="level1"><div class="li"> washed 2x 5 min with EtOH in slideholder</div>
</li>
<li class="level1"><div class="li"> filled slideholder again with EtOH and discarded it immediately</div>
</li>
<li class="level1"><div class="li"> dried with wafergun</div>
</li>
<li class="level1"><div class="li"> spots of DNA were put on flow cells according to the following pattern</div>
</li>
<li class="level1"><div class="li"> incubation overnight at room temperature in humid atmosphere</div>
</li>
</ul>
<p>
spotting pattern (slide 60a):
<a class="media" href="https://static.igem.org/mediawiki/2015/4/43/Freiburg_labjournal-surchem-spotting_pattern_3flowchamber.png" title="labjournal:surchem:spotting_pattern_3flowchamber.png"><img align="left" alt="" class="medialeft" src="https://static.igem.org/mediawiki/2015/4/43/Freiburg_labjournal-surchem-spotting_pattern_3flowchamber.png" width="200"/></a>
</p>
<div class="table sectionedit53"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> sample </th><th class="col2"> concentration </th><th class="col3"> spot size </th>
</tr>
<tr class="row1">
<td class="col0">1</td><td class="col1">HA-GFP-Halo-His-His (Tobi)</td><td class="col2">25 ng/µl</td><td class="col3">1 µl</td>
</tr>
<tr class="row2">
<td class="col0">2</td><td class="col1">Cy5-HA-GFP-His-His</td><td class="col2">12.5 ng/µl</td><td class="col3">3 µl</td>
</tr>
<tr class="row3">
<td class="col0">3</td><td class="col1">Cy5-HA-GFP-His-His</td><td class="col2">44.8 ng/µl</td><td class="col3">1 µl</td>
</tr>
<tr class="row4">
<td class="col0">4</td><td class="col1">Cy5-HA-GFP-His-His</td><td class="col2">28.7 ng/µl</td><td class="col3">0.5 µl</td>
</tr>
<tr class="row5">
<td class="col0">5</td><td class="col1">HA-GFP-Halo-His-His (Tobi)</td><td class="col2">50 ng/µl</td><td class="col3">1 µl</td>
</tr>
<tr class="row6">
<td class="col0">6</td><td class="col1">Cy5-HA-GFP-His-His</td><td class="col2">17.9 ng/µl</td><td class="col3">2 µl</td>
</tr>
<tr class="row7">
<td class="col0">7</td><td class="col1">Cy5-HA-GFP-His-His</td><td class="col2">22.4 ng/µl</td><td class="col3">3 µl</td>
</tr>
<tr class="row8">
<td class="col0">8</td><td class="col1">Cy5-HA-GFP-His-His</td><td class="col2">44.8 ng/µl</td><td class="col3">0.2 µl</td>
</tr>
<tr class="row9">
<td class="col0">9</td><td class="col1">HA-GFP-Halo-His-His (Tobi)</td><td class="col2">99 ng/µl</td><td class="col3">1 µl</td>
</tr>
</table></div>
<!-- EDIT53 TABLE [18986-19419] -->
<p>
spotting pattern (slide 60b):
<a class="media" href="https://static.igem.org/mediawiki/2015/a/a9/Freiburg_labjournal-surchem-spotting_pattern_60b.png" title="labjournal:surchem:spotting_pattern_60b.png"><img align="left" alt="" class="medialeft" src="https://static.igem.org/mediawiki/2015/a/a9/Freiburg_labjournal-surchem-spotting_pattern_60b.png" width="200"/></a>
</p>
<div class="table sectionedit54"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> sample </th><th class="col2"> concentration </th><th class="col3"> spot size </th>
</tr>
<tr class="row1">
<td class="col0">1</td><td class="col1">HA-GFP-Halo-His-His (Tobi)</td><td class="col2">99 ng/µl</td><td class="col3">3 µl</td>
</tr>
<tr class="row2">
<td class="col0">2</td><td class="col1">HA-GFP-Halo-His-His (Tobi)</td><td class="col2">50 ng/µl</td><td class="col3">3 µl</td>
</tr>
<tr class="row3">
<td class="col0">3</td><td class="col1">PCR-Mix von Cy5-HA-GFP-His-His (44.8 ng/µl)</td><td class="col2">-</td><td class="col3">14 µl</td>
</tr>
<tr class="row4">
<td class="col0">4</td><td class="col1">HA-GFP-Halo-His-His (Tobi)</td><td class="col2">25 ng/µl</td><td class="col3">3 µl</td>
</tr>
<tr class="row5">
<td class="col0">5</td><td class="col1">Cy5-HA-GFP-His-His</td><td class="col2">22.4 ng/µl</td><td class="col3">6 µl</td>
</tr>
</table></div>
<!-- EDIT54 TABLE [19506-19788] -->
</div>
<!-- EDIT52 SECTION "2015.08.24" [18278-19789] -->
<h3 class="sectionedit55">2015.08.25</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were let dry in at 60°C for 30 min still in humid atmosphere</div>
</li>
<li class="level1"><div class="li"> expression was effected with 15 µl (60b)/7 µl (60a each flow cell) of Koko mix for 2h at 37°C</div>
</li>
<li class="level1"><div class="li"> flow cells were then checked for fluorescence with microscope of AG Eimer</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> no fluorescence could be observed</div>
</li>
</ul>
<p>
<p><div class="noteimportant">Images are upside-down, meaning the upper line is line 3 (spots 7-9)
</div></p>
</p>
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/7/71/Freiburg_labjournal-surchem-igem_20150825_60a.png" title="labjournal:surchem:igem_20150825_60a.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/71/Freiburg_labjournal-surchem-igem_20150825_60a.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/7/71/Freiburg_labjournal-surchem-igem_20150825_60a.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>60a</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/e/e3/Freiburg_labjournal-surchem-igem_20150825_60b.png" title="labjournal:surchem:igem_20150825_60b.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/e/e3/Freiburg_labjournal-surchem-igem_20150825_60b.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/e/e3/Freiburg_labjournal-surchem-igem_20150825_60b.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>60b</div></div></div>
</div>
<!-- EDIT55 SECTION "2015.08.25" [19790-20349] -->
<h3 class="sectionedit56">2015.08.26</h3>
<div class="level3">
</div>
<h4>Results</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> after storage at 4°C in humid atmosphere overnight flow cells were again checked for fluorescence with microscope of AG Eimer</div>
</li>
<li class="level1"><div class="li"> no fluorescence could be observed</div>
</li>
</ul>
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/2/23/Freiburg_labjournal-surchem-igem_20150826_60a.png" title="labjournal:surchem:igem_20150826_60a.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/23/Freiburg_labjournal-surchem-igem_20150826_60a.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/2/23/Freiburg_labjournal-surchem-igem_20150826_60a.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>60a</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/d/d1/Freiburg_labjournal-surchem-igem_20150826_60b.png" title="labjournal:surchem:igem_20150826_60b.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/d/d1/Freiburg_labjournal-surchem-igem_20150826_60b.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/d/d1/Freiburg_labjournal-surchem-igem_20150826_60b.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>60b</div></div></div>
</div>
<!-- EDIT56 SECTION "2015.08.26" [20350-20669] -->
<h2 class="sectionedit57">Experiment 59d on slide cellfree expressed GFP on Ni-NTA slides for iRIf</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT57 SECTION "Experiment 59d on slide cellfree expressed GFP on Ni-NTA slides for iRIf" [20670-20876] -->
<h3 class="sectionedit58">2015.08.21</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 3 PDITC slides</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 l/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 3 h</div>
</li>
<li class="level1"><div class="li"> </div>
</li>
</ul>
</div>
<!-- EDIT58 SECTION "2015.08.21" [20877-21049] -->
<h3 class="sectionedit59">2015.08.25</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slides were sandwiched with NTA-solution and incubated o/n at 4 °C</div>
</li>
</ul>
</div>
<!-- EDIT59 SECTION "2015.08.25" [21050-21170] -->
<h3 class="sectionedit60">2015.08.26</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slides were blocked with APTES blocking solution for 1 h</div>
</li>
<li class="level1"><div class="li"> slides were washed 2x with PBS</div>
</li>
<li class="level1"><div class="li"> slides were incubated with NiSO<sub>4</sub>-solution for 1 h and 20 min</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS/2x diluted PBS</div>
</li>
</ul>
<ul>
<li class="level1"><div class="li"> cell free GFP Expression mix was spotted on slides (15 µL per spot)and incubated it 3 h at 37 °C</div>
</li>
<li class="level1"><div class="li"> spotted off- slide cell free expressed GFP on the two slides after the first incubation and incubated o/n at 4°C </div>
</li>
</ul>
<p>
spotting pattern:
<img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:spotting_pattern_9.png" width="400"/>
</p>
<p>
slide-#: 303,466
</p>
<div class="table sectionedit61"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1-3 </td><td class="col1"> MM + DNA</td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0"> 4-5 </td><td class="col1"> MM - DNA) </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0"> 6 </td><td class="col1"> GFP (off-slide cell free expressed) </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0"> 7 </td><td class="col1 leftalign"> His-GFP desalt.  </td><td class="col2"> ~1 mg/mL </td>
</tr>
<tr class="row5">
<td class="col0"> 8 </td><td class="col1"> bBSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
</table></div>
<!-- EDIT61 TABLE [21758-21938] -->
</div>
<!-- EDIT60 SECTION "2015.08.26" [21171-21939] -->
<h3 class="sectionedit62">2015.08.27</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocked spots 2x with 10 mg/mL BSA in PBS for 5 min</div>
</li>
<li class="level1"><div class="li"> blocked slides with 10 mg/mL BSA in PBS for 30 min</div>
</li>
<li class="level1"><div class="li"> washed slides with PBS/diluted PBS + 20 mM imidazole/diluted PBS</div>
</li>
</ul>
<p>
Flush protocol:
slide 466 and 303
</p>
<div class="table sectionedit63"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">450</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">450</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">450</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Anti-GFP (goat, biotinylated)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">StrepCy5</td><td class="col1">6</td><td class="col2">30</td><td class="col3">300</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT63 TABLE [22204-22459] -->
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/63/Freiburg_labjournal-surchem-exp_59d.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_59d.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>ROI selection of slide 466</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/1/19/Freiburg_labjournal-surchem-exp_59d_scanner.jpg" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_59d_scanner.jpg" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>picture of slide 466 from the microarray scanner (exitation at 635 nm</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/5/51/Freiburg_labjournal-surchem-exp_59d_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_59d_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>binding curves of slide 466</div></div></div>
<p>
→ salt deposits on the surface of slide 303 made an evaluation difficult
</p>
</div>
<!-- EDIT62 SECTION "2015.08.27" [21940-22839] -->
<h2 class="sectionedit64">Experiment 59c PDITC/Ni-NTA slides for iRIf - cell free Expression on slides</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT64 SECTION "Experiment 59c PDITC/Ni-NTA slides for iRIf - cell free Expression on slides" [22840-23050] -->
<h3 class="sectionedit65">2015.08.21</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 3 PDITC slides</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 l/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 3 h</div>
</li>
</ul>
</div>
<!-- EDIT65 SECTION "2015.08.21" [23051-23218] -->
<h3 class="sectionedit66">2015.08.23</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 3 Ni-NTA slides according to <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation protocol</a></div>
</li>
<li class="level1"><div class="li"> PDITC slides were incubated o/n with NTA at 4°C</div>
</li>
<li class="level1"><div class="li"> slides were blocked in APTES blocking solution for 30 min</div>
</li>
<li class="level1"><div class="li"> slides were incubated in NiSO4 for 1 h</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS/diluted PBS</div>
</li>
<li class="level1"><div class="li"> slides were stored at 4 °C</div>
</li>
<li class="level1"><div class="li"> </div>
</li>
</ul>
</div>
<!-- EDIT66 SECTION "2015.08.23" [23219-23607] -->
<h3 class="sectionedit67">2015.08.25</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spotted free expression mix on two slides (<strong>429</strong>, <strong>424</strong>) and incubated it 3 h at 37 °C</div>
</li>
<li class="level1"><div class="li"> spotting mask was accidently removed after incubation –&gt; spotted Solutions probably mixed (no definite spots)</div>
</li>
<li class="level1"><div class="li"> spotted off- slide cell free expressed GFP on the two slides after the first incubation and incubated o/n at 4°C </div>
</li>
<li class="level1"><div class="li"> spotted 1 slide (<strong>500</strong>) with only off-slide cell-free expressed GFP and incubated o/n at 4°C </div>
</li>
</ul>
<p>
spotting pattern:
<img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:spotting_pattern_9.png" width="400"/>
</p>
<p>
slide-#: <strong>429, 424</strong>
</p>
<div class="table sectionedit68"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1-3 </td><td class="col1"> MM + DNA</td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0"> 4-5 </td><td class="col1"> MM - DNA) </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0"> 6 </td><td class="col1"> GFP (off-slide cell free expressed) </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0"> 7 </td><td class="col1 leftalign"> His-GFP desalt.  </td><td class="col2"> ~1 mg/mL </td>
</tr>
<tr class="row5">
<td class="col0"> 8 </td><td class="col1"> bBSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
</table></div>
<!-- EDIT68 TABLE [24188-24368] -->
<p>
spotting pattern:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<p>
slide-#: <strong>500</strong>
</p>
<div class="table sectionedit69"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1-3 </td><td class="col1"> MM + DNA</td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0"> 4-6 </td><td class="col1"> MM - DNA </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0"> 7-8</td><td class="col1"> MM + DNA (off-slide cell free expressed, 15 µl reachtion ) </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0"> 10 </td><td class="col1 leftalign"> His-GFP desalt.  </td><td class="col2"> ~1 mg/mL </td>
</tr>
<tr class="row5">
<td class="col0"> 11 </td><td class="col1"> bBSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
</table></div>
<!-- EDIT69 TABLE [24469-24675] -->
</div>
<!-- EDIT67 SECTION "2015.08.25" [23608-24677] -->
<h3 class="sectionedit70">2015.08.26</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked with 10 mg/mL BSA for 30 min</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS/diluted PBS + 20 mM imidazole/diluted PBS</div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
slide 424 and 500
</p>
<div class="table sectionedit71"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">450</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">450</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">450</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Anti-GFP (goat, biotinylated)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">StrepCy5</td><td class="col1">6</td><td class="col2">30</td><td class="col3">300</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT71 TABLE [24948-25203] -->
<p>
429 was not measured because the other slides showed that GFP was expressed, so the Experiment will be repeated and the above mentioned mistake (removal of spotting masks after 37 °C incubation) will be avoided.
</p>
</div>
<h4>Results</h4>
<div class="level4">
<p>
Only slide 500 was evaluated. Slide 424 and 429 showed very bad surface during measurements which was wiped off → Difficult to evaluate
Slide 500: spots 4 and 8 had to be excluded from evaluation.
</p>
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/2b/Freiburg_labjournal-roi_selection_20150826_500.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:roi_selection_20150826_500.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 500: ROI selection</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/e/e3/Freiburg_labjournal-binding_curve_20150826_500.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:binding_curve_20150826_500.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 500: Binding curves</div></div></div>
</div>
<!-- EDIT70 SECTION "2015.08.26" [24678-25804] -->
<h2 class="sectionedit72">Experiment 59b PDITC/Ni-NTA slides for iRIf - cell free expressed Tetanus</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 PDITC slides (208, 216)</div>
</li>
</ul>
</div>
<!-- EDIT72 SECTION "Experiment 59b PDITC/Ni-NTA slides for iRIf - cell free expressed Tetanus" [25805-26074] -->
<h3 class="sectionedit73">2015.08.21</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 2 PDITC slides</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 l/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 3 h</div>
</li>
</ul>
</div>
<!-- EDIT73 SECTION "2015.08.21" [26075-26243] -->
<h3 class="sectionedit74">2015.08.23</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 2 Ni-NTA slides according to <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation protocol</a></div>
</li>
<li class="level1"><div class="li"> PDITC slides were incubated o/n with NTA at 4°C</div>
</li>
<li class="level1"><div class="li"> slides were blocked in APTES blocking solution for 30 min</div>
</li>
<li class="level1"><div class="li"> slides were incubated in NiSO4 for 1 h</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS/diluted PBS</div>
</li>
<li class="level1"><div class="li"> slides were stored at 4 °C</div>
</li>
</ul>
</div>
<!-- EDIT74 SECTION "2015.08.23" [26244-26629] -->
<h3 class="sectionedit75">2015.08.25</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spotted with cell-free expressed Tetanus samples, GFP and bBSA</div>
</li>
</ul>
<p>
spotting pattern:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<p>
slide-#: 216
</p>
<div class="table sectionedit76"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1 </td><td class="col1"> B+ (1) </td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0"> 2 </td><td class="col1"> B+ (2) </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0"> 3 </td><td class="col1"> B+ (3) </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0"> 4 </td><td class="col1"> B- (1) </td><td class="col2"> </td>
</tr>
<tr class="row5">
<td class="col0"> 5 </td><td class="col1"> B- (2) </td><td class="col2"> </td>
</tr>
<tr class="row6">
<td class="col0"> 6 </td><td class="col1"> K+ (1) </td><td class="col2"> </td>
</tr>
<tr class="row7">
<td class="col0"> 7 </td><td class="col1"> His-GFP desalt. </td><td class="col2"> ~0.5 mg/mL </td>
</tr>
<tr class="row8">
<td class="col0"> 8 </td><td class="col1"> His-GFP lysate</td><td class="col2"> </td>
</tr>
<tr class="row9">
<td class="col0"> 9 </td><td class="col1"> bBSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
<tr class="row10">
<td class="col0"> 10 </td><td class="col1"> K- </td><td class="col2"> </td>
</tr>
<tr class="row11">
<td class="col0"> 11 </td><td class="col1"> BSA </td><td class="col2"> </td>
</tr>
</table></div>
<!-- EDIT76 TABLE [26842-27092] -->
<p>
slide-#: 208
</p>
<div class="table sectionedit77"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1 </td><td class="col1"> K+ (1) </td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0"> 2 </td><td class="col1"> K+ (2) </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0"> 3 </td><td class="col1"> K+ (3) </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0"> 4 </td><td class="col1"> K- (1) </td><td class="col2"> </td>
</tr>
<tr class="row5">
<td class="col0"> 5 </td><td class="col1"> K- (2) </td><td class="col2"> </td>
</tr>
<tr class="row6">
<td class="col0"> 6 </td><td class="col1"> B+ (1) </td><td class="col2"> </td>
</tr>
<tr class="row7">
<td class="col0"> 7 </td><td class="col1"> His-GFP desalt. </td><td class="col2"> ~0.5 mg/mL </td>
</tr>
<tr class="row8">
<td class="col0"> 8 </td><td class="col1"> His-GFP lysate</td><td class="col2"> </td>
</tr>
<tr class="row9">
<td class="col0"> 9 </td><td class="col1"> bBSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
<tr class="row10">
<td class="col0"> 10 </td><td class="col1"> B- </td><td class="col2"> </td>
</tr>
<tr class="row11">
<td class="col0"> 11 </td><td class="col1"> BSA </td><td class="col2"> </td>
</tr>
</table></div>
<!-- EDIT77 TABLE [27107-27357] -->
</div>
<!-- EDIT75 SECTION "2015.08.25" [26630-27358] -->
<h3 class="sectionedit78">2015.08.26</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked with 10 mg/mL BSA for 30 min</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS/diluted PBS + 20 mM imidazole/diluted PBS</div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<div class="table sectionedit79"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer (PBS)</td><td class="col1">1</td><td class="col2">60</td><td class="col3">450</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0 leftalign">BSA	</td><td class="col1">2</td><td class="col2">60</td><td class="col3">450</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer	(PBS)</td><td class="col1">3</td><td class="col2">60</td><td class="col3">450</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Anti-Tetanus (goat, pk)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">25 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer	(PBS)</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">Anti-GFP (biotinylated, Goat)</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer	(PBS)</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">Anti-Goat</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer	(PBS)</td><td class="col1">9</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row10">
<td class="col0">StrepCy5</td><td class="col1">10</td><td class="col2">30</td><td class="col3">300</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row11">
<td class="col0">Buffer	(PBS)</td><td class="col1">11</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT79 TABLE [27610-28019] -->
</div>
<h4>Results</h4>
<div class="level4">
<p>
Slide 208: Only 5 spots (spot 3, 5, 7, 8 and 9) could be evaluated.
</p>
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/d/df/Freiburg_labjournal-roi_selection_20150826_208.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:roi_selection_20150826_208.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 208: ROI selection</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/3/3e/Freiburg_labjournal-roi_selection_20150826_216.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:roi_selection_20150826_216.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 216: ROI selection</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/a/a4/Freiburg_labjournal-binding_curve_20150826_208.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:binding_curve_20150826_208.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 208: Binding curves</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/b/b9/Freiburg_labjournal-binding_curve_20150826_216.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:binding_curve_20150826_216.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 216: Binding curves</div></div></div>
</div>
<!-- EDIT78 SECTION "2015.08.26" [27359-28447] -->
<h2 class="sectionedit80">Experiment 59a: PDITC/Ni-NTA slides for iRIf - Tetanus with own blood</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 PDITC slides</div>
</li>
</ul>
</div>
<!-- EDIT80 SECTION "Experiment 59a: PDITC/Ni-NTA slides for iRIf - Tetanus with own blood" [28448-28703] -->
<h3 class="sectionedit81">2015.08.21</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 2 PDITC slides </div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 l/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 3 h</div>
</li>
<li class="level1"><div class="li"> spotted 3 µL of Antigen and controls (GFP) </div>
</li>
</ul>
<p>
spotting pattern:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<p>
slide-#: 287, 504
</p>
<div class="table sectionedit82"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1-2 </td><td class="col1"> Tetanus (11) desalt </td><td class="col2"> 460 µg/mL </td>
</tr>
<tr class="row2">
<td class="col0"> 3-4 </td><td class="col1"> Tetanus (11) desalt </td><td class="col2"> 100 µg/mL </td>
</tr>
<tr class="row3">
<td class="col0"> 5-6 </td><td class="col1"> Tetanus (11) desalt </td><td class="col2"> 20 µg/mL </td>
</tr>
<tr class="row4">
<td class="col0"> 7-8 </td><td class="col1"> His-GFP desalt. </td><td class="col2"> ~0.5 mg/mL </td>
</tr>
<tr class="row5">
<td class="col0"> 9 </td><td class="col1"> bBSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
<tr class="row6">
<td class="col0"> 10-11 </td><td class="col1"> Tetanus (11) lysate </td><td class="col2"> </td>
</tr>
</table></div>
<!-- EDIT82 TABLE [29023-29282] -->
</div>
<!-- EDIT81 SECTION "2015.08.21" [28704-29283] -->
<h3 class="sectionedit83">2015.08.22</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA in PBS for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked with 10 mg/mL BSA in PBS for 30 min</div>
</li>
<li class="level1"><div class="li"> slides were washed with Aqua dest and dried</div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
Flush protocol slide 287
</p>
<div class="table sectionedit84"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer (PBS)</td><td class="col1">1</td><td class="col2">60</td><td class="col3">720</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0 leftalign">BSA	</td><td class="col1">2</td><td class="col2">60</td><td class="col3">900</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer	(PBS)</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Serum rabbit Anti-GFP</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1:330</td>
</tr>
<tr class="row5">
<td class="col0">Buffer	(PBS)</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">Serum (+) SIG002</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1:10</td>
</tr>
<tr class="row7">
<td class="col0">Buffer	(PBS)</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">Anti-Human</td><td class="col1">8</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer	(PBS)</td><td class="col1">9</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT84 TABLE [29543-29875] -->
<p>
no Strep-Cy5 was flushed over slide –&gt; no scanner Pictures
</p>
<p>
Flush protocol slide 504
</p>
<div class="table sectionedit85"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer (PBS)</td><td class="col1">1</td><td class="col2">60</td><td class="col3">720</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0 leftalign">BSA	</td><td class="col1">2</td><td class="col2">60</td><td class="col3">900</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer	(PBS)</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Serum (-) SIG002</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1:10</td>
</tr>
<tr class="row5">
<td class="col0">Buffer	(PBS)</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">Serum (+) SIG002</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1:10</td>
</tr>
<tr class="row7">
<td class="col0">Buffer	(PBS)</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">Anti-Human</td><td class="col1">8</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer	(PBS)</td><td class="col1">9</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row10">
<td class="col0">Anti-GFP (biotinylated, Goat)</td><td class="col1">10</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1 ug/ml</td>
</tr>
<tr class="row11">
<td class="col0">Buffer	(PBS)</td><td class="col1">11</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row12">
<td class="col0">Strep-Cy5</td><td class="col1">12</td><td class="col2">60</td><td class="col3">300</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row13">
<td class="col0">Buffer	(PBS)</td><td class="col1">13</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT85 TABLE [29963-30425] -->
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/77/Freiburg_labjournal-surchem-exp_59a_slide287.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_59a_slide287.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 287: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/27/Freiburg_labjournal-surchem-exp_59a_slide287_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_59a_slide287_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 287: Binding curves</div></div></div><hr/>
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/65/Freiburg_labjournal-surchem-exp_59a_slide504.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_59a_slide504.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 504: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/a/ae/Freiburg_labjournal-surchem-exp_59a_slide504_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_59a_slide504_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 504: Binding curves</div></div></div>
<p>
→ detection of Tetanus antibodys from blood serum after vaccination is possible<br/>

→ huge cross reactivity with bBSA spot→probably due to biodinidase enzyme
</p>
</div>
<!-- EDIT83 SECTION "2015.08.22" [29284-30963] -->
<h2 class="sectionedit86">Experiment 58: ELISA with Tetanus-Toxin and blood</h2>
<div class="level2">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> Check if antibodies in blood of freshly vaccinated Patient SIG002 binds to our self-expressed Tetanus Antigen</div>
</li>
<li class="level1"><div class="li"> detection of antibodies in blood with anti-human (mouse)</div>
</li>
<li class="level1"><div class="li"> detection of anti-human with HRP-labeled anti-mouse</div>
</li>
</ul>
</div>
<!-- EDIT86 SECTION "Experiment 58: ELISA with Tetanus-Toxin and blood" [30964-31280] -->
<h3 class="sectionedit87">2015.08.20</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 100 µL of each Tet-Tox and bBSA concentration were pipetted into maxisorb-plate</div>
</li>
</ul>
<div class="table sectionedit88"><table class="inline">
<tr class="row0">
<th class="col0"> </th><th class="col1"> 1 </th><th class="col2"> 2 </th><th class="col3"> 3 </th>
</tr>
<tr class="row1">
<td class="col0">A</td><td class="col1"> Tet-Tox 100 µg/mL </td><td class="col2"> bBSA 100 µg/mL </td><td class="col3"> bBSA 0.4 µg/mL </td>
</tr>
<tr class="row2">
<td class="col0">B</td><td class="col1"> Tet-Tox 50 µg/mL </td><td class="col2"> bBSA 50 µg/mL </td><td class="col3"> bBSA 0.2 µg/mL </td>
</tr>
<tr class="row3">
<td class="col0">C</td><td class="col1"> Tet-Tox 25 µg/mL </td><td class="col2"> bBSA 25 µg/mL </td><td class="col3"> bBSA 0.1 µg/mL </td>
</tr>
<tr class="row4">
<td class="col0">D</td><td class="col1"> Tet-Tox 12 µg/mL </td><td class="col2"> bBSA 12 µg/mL </td><td class="col3"> bBSA 0.05 µg/mL </td>
</tr>
<tr class="row5">
<td class="col0">E</td><td class="col1"> Tet-Tox 6 µg/mL </td><td class="col2"> bBSA 6 µg/mL </td><td class="col3"> bBSA 0.025 µg/mL </td>
</tr>
<tr class="row6">
<td class="col0">F</td><td class="col1"> Tet-Tox 3 µg/mL </td><td class="col2"> bBSA 3 µg/mL </td><td class="col3"> bBSA 0.012 µg/mL </td>
</tr>
<tr class="row7">
<td class="col0">G</td><td class="col1"> Tet-Tox 1.5 µg/mL </td><td class="col2"> bBSA 1.5 µg/mL </td><td class="col3"> bBSA 0.006 µg/mL </td>
</tr>
<tr class="row8">
<td class="col0">H</td><td class="col1"> neg (PBS) </td><td class="col2"> bBSA 0.8 µg/mL </td><td class="col3"> neg (PBS) </td>
</tr>
</table></div>
<!-- EDIT88 TABLE [31414-31895] -->
</div>
<!-- EDIT87 SECTION "2015.08.20" [31281-31896] -->
<h3 class="sectionedit89">2015.08.21</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 8:34 am: all wells exept H1 were blocked with 5 mg/mL BSA in PBS</div>
</li>
<li class="level1"><div class="li"> wells were washed with PBS 2 times for ~2 min</div>
</li>
<li class="level1"><div class="li"> 10:00 am: 100 µL of 1/10 diluted blood Serum were added to well A1-H1 and again 100 µL of 5 mg/mL BSA in PBS to A2-H2 and A3-H3</div>
</li>
<li class="level1"><div class="li"> all solutions were overlayed with 100 µL PBS</div>
</li>
<li class="level1"><div class="li"> wells were washed with PBS 3 times for ~2 min</div>
</li>
<li class="level1"><div class="li"> 11.07 am: 100 µL of anti-human (1 µg/mL) were added to well A1-H1 and again 100 µL of 5 mg/mL BSA in PBS to A2-H2 and A3-H3</div>
</li>
<li class="level1"><div class="li"> all solutions were overlayed with 100 µL PBS</div>
</li>
<li class="level1"><div class="li"> wells were washed with PBS 3 times for ~2 min</div>
</li>
<li class="level1"><div class="li"> 1:00 pm: 100 µL of HRP-anti-mouse (1 µg/mL) were added to well A1-H1 and 100 µL of HRP-Strep (0.5 µg/mL) to A2-H2 and A3-H3</div>
</li>
<li class="level1"><div class="li"> all solutions were overlayed with 100 µL PBS</div>
</li>
<li class="level1"><div class="li"> wells were washed with PBS 5 times for ~2 min</div>
</li>
</ul>
<ul>
<li class="level1"><div class="li"> Hydrogenperoxide and 3,3´,5,5´-Tetramethylbenzidine were added to each well and the Absorption was measured in the plate reader</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 trien" style="width:610px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/8/8e/Freiburg_labjournal-surchem-20150821_elisa_evaluation.png" title="labjournal:surchem:20150821_elisa_evaluation.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/8/8e/Freiburg_labjournal-surchem-20150821_elisa_evaluation.png" width="600"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/8/8e/Freiburg_labjournal-surchem-20150821_elisa_evaluation.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Absorbance over time for Tetanus-ELISA Test with own blood</div></div></div>
</div>
<!-- EDIT89 SECTION "2015.08.21" [31897-33031] -->
<h2 class="sectionedit90">Experiment 57d: Ni NTA iRIf slides - Salmonella</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT90 SECTION "Experiment 57d: Ni NTA iRIf slides - Salmonella" [33032-33214] -->
<h3 class="sectionedit91">2015.08.18</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 PDITC iRif slides</div>
</li>
<li class="level1"><div class="li"> slide#:</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slides were washed according to <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Slide_Wash" title="protocols:slide_wash">iRIf slide wash steps</a></div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation 3.5 h</div>
</li>
</ul>
</div>
<!-- EDIT91 SECTION "2015.08.18" [33215-33505] -->
<h3 class="sectionedit92">2015.08.20</h3>
<div class="level3">
<ul>
<li class="level1"><div class="li"> slides were blocked in APTES blocking solution for 45 min</div>
</li>
<li class="level1"><div class="li"> slides were incubated in NiSO<sub>4</sub> for 1 h</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS/diluted PBS</div>
</li>
<li class="level1"><div class="li"> slides were stored at 4 °C</div>
</li>
</ul>
</div>
<!-- EDIT92 SECTION "2015.08.20" [33506-33719] -->
<h3 class="sectionedit93">2015.08.21</h3>
<div class="level3">
<p>
* spotted 3 µL of Salmonella desalt and lysate and controls (GFP-Lysate, GFP-desalt, bBSA) 
</p>
<p>
spotting pattern:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<p>
slide-#: 012, 426
</p>
<div class="table sectionedit94"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1-3 </td><td class="col1"> Salmonella (15) desalt </td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0"> 4-6 </td><td class="col1"> Salmonella (15) lysate </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0"> 7 </td><td class="col1"> His-GFP desalt. </td><td class="col2"> 0.2 mg/ml </td>
</tr>
<tr class="row4">
<td class="col0"> 8 </td><td class="col1"> His-GFP Lysate </td><td class="col2"> ~ 0.5 mg/ml </td>
</tr>
<tr class="row5">
<td class="col0"> 9 </td><td class="col1"> His-GFP Lysate </td><td class="col2"> ~ 0.5 mg/ml </td>
</tr>
<tr class="row6">
<td class="col0"> 10 </td><td class="col1"> nt-GFP Lysate </td><td class="col2"> ~ 0.5 mg/ml </td>
</tr>
<tr class="row7">
<td class="col0"> 11 </td><td class="col1"> bBSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
</table></div>
<!-- EDIT94 TABLE [33935-34210] -->
</div>
<!-- EDIT93 SECTION "2015.08.21" [33720-34211] -->
<h3 class="sectionedit95">2015.08.22</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA in <strong>TBS</strong> for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked with 10 mg/mL BSA in <strong>TBS</strong> for 30 min</div>
</li>
<li class="level1"><div class="li"> slides were washed with <strong>TBS</strong>/diluted <strong>TBS</strong> + 20 mM imidazole/diluted <strong>TBS</strong></div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<div class="table sectionedit96"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer (TBS)</td><td class="col1">1</td><td class="col2">60</td><td class="col3">720</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">His-HIV-Lysate (17)/BSA </td><td class="col1">2</td><td class="col2">60</td><td class="col3">900</td><td class="col4">1:1</td>
</tr>
<tr class="row3">
<td class="col0">Buffer	(TBS)</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Anti-GFP (biotinylated, goat)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer	(TBS)</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">StrepCy5</td><td class="col1">6</td><td class="col2">60</td><td class="col3">300</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer	(TBS)</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">Anti-Salmonella desalt (13)</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">~60 µg/mL</td>
</tr>
<tr class="row9">
<td class="col0">Buffer	(TBS)</td><td class="col1">9</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row10">
<td class="col0">Anti-Salmonella Lysate (13)</td><td class="col1">10</td><td class="col2">30</td><td class="col3">600</td><td class="col4">~60 µg/mL</td>
</tr>
<tr class="row11">
<td class="col0">Buffer	(TBS)</td><td class="col1">11</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT96 TABLE [34491-34942] -->
</div>
<!-- EDIT95 SECTION "2015.08.22" [34212-34944] -->
<h2 class="sectionedit97">Experiment 57c: Ni NTA iRIf slides - cell free expressed GFP</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT97 SECTION "Experiment 57c: Ni NTA iRIf slides - cell free expressed GFP" [34945-35139] -->
<h3 class="sectionedit98">2015.08.18</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 4 PDITC iRif slides</div>
</li>
<li class="level1"><div class="li"> slide#: 423, 215, 254, 024</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slides were washed according to <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Slide_Wash" title="protocols:slide_wash">iRIf slide wash steps</a></div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation 3.5 h</div>
</li>
</ul>
</div>
<!-- EDIT98 SECTION "2015.08.18" [35140-35449] -->
<h3 class="sectionedit99">2015.08.20</h3>
<div class="level3">
<ul>
<li class="level1"><div class="li"> slides were blocked in APTES blocking solution for 45 min</div>
</li>
<li class="level1"><div class="li"> slides were incubated in NiSO<sub>4</sub> for 1 h</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS/diluted PBS</div>
</li>
<li class="level1"><div class="li"> spotted 3 µL of cell free expressed GFPs and controls (GFP-Lysate, GFP-desalt, bBSA) </div>
</li>
</ul>
<p>
spotting pattern:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<p>
slide-#: 423, 254
</p>
<div class="table sectionedit100"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1 </td><td class="col1"> BK HA-GFP-His6His6 (E2) </td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0"> 2 </td><td class="col1"> KK HA-GFP-His6His6 (G2) </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0"> 3 </td><td class="col1"> PP HA-GFP-His6His6 (I2) </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0"> 4 </td><td class="col1"> BP HA-GFP-His6His6 (K2) </td><td class="col2"> </td>
</tr>
<tr class="row5">
<td class="col0"> 5 </td><td class="col1"> BK His10-GFP-Spy (E3) </td><td class="col2"> </td>
</tr>
<tr class="row6">
<td class="col0"> 6 </td><td class="col1"> KK His10-GFP-Spy (G3) </td><td class="col2"> </td>
</tr>
<tr class="row7">
<td class="col0"> 7 </td><td class="col1"> His-GFP desalt. </td><td class="col2"> 0.2 mg/ml </td>
</tr>
<tr class="row8">
<td class="col0"> 8 </td><td class="col1"> His-GFP Lysate </td><td class="col2"> ~ 0.5 mg/ml </td>
</tr>
<tr class="row9">
<td class="col0"> 9 </td><td class="col1"> bBSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
<tr class="row10">
<td class="col0"> 10 </td><td class="col1"> PP His10-GFP-Spy (I3) </td><td class="col2"> </td>
</tr>
<tr class="row11">
<td class="col0"> 11 </td><td class="col1"> BP His10-GFP-Spy (K3) </td><td class="col2"> </td>
</tr>
</table></div>
<!-- EDIT100 TABLE [35823-36219] -->
<p>
slide-#: 024, 215
</p>
<div class="table sectionedit101"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1 </td><td class="col1"> BK HA-GFP-His6His6 (F2) </td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0"> 2 </td><td class="col1"> KK HA-GFP-His6His6 (H2) </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0"> 3 </td><td class="col1"> PP HA-GFP-His6His6 (J2) </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0"> 4 </td><td class="col1"> BP HA-GFP-His6His6 (L2) </td><td class="col2"> </td>
</tr>
<tr class="row5">
<td class="col0"> 5 </td><td class="col1"> BK His10-GFP-Spy (F3) </td><td class="col2"> </td>
</tr>
<tr class="row6">
<td class="col0"> 6 </td><td class="col1"> KK His10-GFP-Spy (H3) </td><td class="col2"> </td>
</tr>
<tr class="row7">
<td class="col0"> 7 </td><td class="col1"> His-GFP desalt. </td><td class="col2"> 0.2 mg/ml </td>
</tr>
<tr class="row8">
<td class="col0"> 8 </td><td class="col1"> His-GFP Lysate </td><td class="col2"> ~ 0.5 mg/ml </td>
</tr>
<tr class="row9">
<td class="col0"> 9 </td><td class="col1"> bBSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
<tr class="row10">
<td class="col0"> 10 </td><td class="col1"> PP His10-GFP-Spy (J3) </td><td class="col2"> </td>
</tr>
<tr class="row11">
<td class="col0"> 11 </td><td class="col1"> BP His10-GFP-Spy (L3) </td><td class="col2"> </td>
</tr>
</table></div>
<!-- EDIT101 TABLE [36240-36636] -->
</div>
<!-- EDIT99 SECTION "2015.08.20" [35450-36637] -->
<h3 class="sectionedit102">2015.08.21</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked with 10 mg/mL BSA for 30 min</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS/diluted PBS + 20 mM Imidazole/diluted PBS</div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
slide 215: old Setup
</p>
<div class="table sectionedit103"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer (PBS)</td><td class="col1">1</td><td class="col2">60</td><td class="col3">720</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0 leftalign">BSA	</td><td class="col1">2</td><td class="col2">60</td><td class="col3">720</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer	(PBS)</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Anti GFP (biotinylated, goat)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer	(PBS)</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">StrepCy5</td><td class="col1">6</td><td class="col2">60</td><td class="col3">300</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer	(PBS)</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT103 TABLE [36911-37191] -->
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/8/87/Freiburg_labjournal-surchem-20150821_slide_024_binding_curve.png" title="labjournal:surchem:20150821_slide_024_binding_curve.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/8/87/Freiburg_labjournal-surchem-20150821_slide_024_binding_curve.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/8/87/Freiburg_labjournal-surchem-20150821_slide_024_binding_curve.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>slide_024_binding_curve</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/d/d7/Freiburg_labjournal-surchem-20150821_slide_024_roi_024.png" title="labjournal:surchem:20150821_slide_024_roi_024.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/d/d7/Freiburg_labjournal-surchem-20150821_slide_024_roi_024.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/d/d7/Freiburg_labjournal-surchem-20150821_slide_024_roi_024.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>slide_024_roi_024</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/f/f5/Freiburg_labjournal-surchem-20150821_slide_215_binding_curves.png" title="labjournal:surchem:20150821_slide_215_binding_curves.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/f5/Freiburg_labjournal-surchem-20150821_slide_215_binding_curves.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/f/f5/Freiburg_labjournal-surchem-20150821_slide_215_binding_curves.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>slide_215_binding_curves</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/e/eb/Freiburg_labjournal-surchem-20150821_slide_215_roi.png" title="labjournal:surchem:20150821_slide_215_roi.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/e/eb/Freiburg_labjournal-surchem-20150821_slide_215_roi.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/e/eb/Freiburg_labjournal-surchem-20150821_slide_215_roi.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>slide_215_roi</div></div></div><hr/>
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/3/36/Freiburg_labjournal-surchem-20150821_slide_254_binding_curves.png" title="labjournal:surchem:20150821_slide_254_binding_curves.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/3/36/Freiburg_labjournal-surchem-20150821_slide_254_binding_curves.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/3/36/Freiburg_labjournal-surchem-20150821_slide_254_binding_curves.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>slide_254_binding_curves</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/5/57/Freiburg_labjournal-surchem-20150821_slide_254_roi.png" title="labjournal:surchem:20150821_slide_254_roi.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/5/57/Freiburg_labjournal-surchem-20150821_slide_254_roi.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/5/57/Freiburg_labjournal-surchem-20150821_slide_254_roi.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>slide_254_roi</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/3/37/Freiburg_labjournal-surchem-20150821_slide_423_binding_curves.png" title="labjournal:surchem:20150821_slide_423_binding_curves.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/3/37/Freiburg_labjournal-surchem-20150821_slide_423_binding_curves.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/3/37/Freiburg_labjournal-surchem-20150821_slide_423_binding_curves.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>slide_423_binding_curves</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/0/0e/Freiburg_labjournal-surchem-20150821_slide_423_roi.png" title="labjournal:surchem:20150821_slide_423_roi.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/0/0e/Freiburg_labjournal-surchem-20150821_slide_423_roi.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/0/0e/Freiburg_labjournal-surchem-20150821_slide_423_roi.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>slide_423_roi</div></div></div>
</div>
<!-- EDIT102 SECTION "2015.08.21" [36638-37878] -->
<h2 class="sectionedit104">Experiment 57b: Ni NTA iRIf slides - cell free Expression on Ni-NTA slides</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT104 SECTION "Experiment 57b: Ni NTA iRIf slides - cell free Expression on Ni-NTA slides" [37879-38087] -->
<h3 class="sectionedit105">2015.08.18</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 PDITC iRif slides</div>
</li>
<li class="level1"><div class="li"> slide#: 466, 253</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slides were washed according to <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Slide_Wash" title="protocols:slide_wash">iRIf slide wash steps</a></div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation 3.5 h</div>
</li>
</ul>
</div>
<!-- EDIT105 SECTION "2015.08.18" [38088-38387] -->
<h3 class="sectionedit106">2015.08.20</h3>
<div class="level3">
<ul>
<li class="level1"><div class="li"> slides were blocked in APTES blocking solution for 45 min</div>
</li>
<li class="level1"><div class="li"> slides were incubated in NiSO<sub>4</sub> for 1 h</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS/diluted PBS</div>
</li>
<li class="level1"><div class="li"> two slides (<strong>466</strong>, <strong>253</strong>) were spotted with 15 µl per spot spottingmasks</div>
</li>
</ul>
<p>
Spotting: 
<img alt="" class="media" src="https://static.igem.org/mediawiki/2015/9/99/Freiburg_labjournal-surchem-spotting_pattern_9.png" width="300"/>
</p>
<div class="table sectionedit107"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1-3 </td><td class="col1">His-GFP (cellfree expressed))</td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0"> 4-6 </td><td class="col1">neg. control (cellfree Expression without DNA)</td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0"> 7 </td><td class="col1"> bBSA </td><td class="col2"> 200 µg/ml </td>
</tr>
<tr class="row4">
<td class="col0"> 8 </td><td class="col1"> His GFP lysate</td><td class="col2 leftalign"> </td>
</tr>
</table></div>
<!-- EDIT107 TABLE [38721-38903] --><ul>
<li class="level1"><div class="li"> slides were covered with glass slides to avoid evaporation</div>
</li>
<li class="level1"><div class="li"> slides were incubated for for 3 h at 37 °C</div>
</li>
<li class="level1"><div class="li"> measurement in iRIf</div>
</li>
</ul>
<div class="table sectionedit108"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer (PBS)</td><td class="col1">1</td><td class="col2">60</td><td class="col3">720</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0 leftalign">BSA	</td><td class="col1">2</td><td class="col2">60</td><td class="col3">720</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer	(PBS)</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Anti GFP (biotinylated, goat)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer	(PBS)</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">StrepCy5</td><td class="col1">6</td><td class="col2">60</td><td class="col3">300</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer	(PBS)</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT108 TABLE [39041-39321] -->
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 tleft" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/7c/Freiburg_labjournal-surchem-exp_57b_slide253.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_57b_slide253.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 253: Quotion picture</div></div></div><div class="thumb2 tleft" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/3/36/Freiburg_labjournal-surchem-exp_57b_slide466.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_57b_slide466.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 466: Quotion picture</div></div></div>
<p>
→ no cell free expressed GFP could be detected<br/>

→ on slide 253 even the spotted GFP lysate was not detectable, this is probably due to repeaded problems with the correct flooding of the flow cell during the measurement with the iRIf setup<br/>

→ the experiment will be repeated with additional spots of „off slide“ cell free expressed GFP 
</p>
</div>
<!-- EDIT106 SECTION "2015.08.20" [38388-39852] -->
<h2 class="sectionedit109">Experiment 57a: PDITC iRIf slides - anti-GFP-Serum measurement</h2>
<div class="level2">
</div>
<!-- EDIT109 SECTION "Experiment 57a: PDITC iRIf slides - anti-GFP-Serum measurement" [39853-39925] -->
<h3 class="sectionedit110">2015.08.18</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 PDITC iRif slides</div>
</li>
<li class="level1"><div class="li"> slide#: 315, 502</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slides were washed according to <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Slide_Wash" title="protocols:slide_wash">iRIf slide wash steps</a></div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation 3.5 h</div>
</li>
<li class="level1"><div class="li"> slides were washed and dried with wafer gun</div>
</li>
<li class="level1"><div class="li"> 3 µL of cell free expressed GFP, GFP Lysate, GFP desalt, bBSA, BSA and mCherry were spotted.</div>
</li>
</ul>
<p>
spotting pattern:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<div class="table sectionedit111"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1-2 </td><td class="col1">His-GFP (cellfree expressed))</td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0"> 3 </td><td class="col1"> His-GFP desalt. </td><td class="col2"> 0.2 mg/ml </td>
</tr>
<tr class="row3">
<td class="col0"> 4-5 </td><td class="col1">neg. control (cellfree Expression without DNA)</td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0"> 6-7 </td><td class="col1"> His-mCherry lysate </td><td class="col2"> 1:10 </td>
</tr>
<tr class="row5">
<td class="col0"> 8-9 </td><td class="col1"> His-GFP lysate </td><td class="col2"> 1:10 </td>
</tr>
<tr class="row6">
<td class="col0"> 10 </td><td class="col1">BSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
<tr class="row7">
<td class="col0"> 11 </td><td class="col1">bBSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
</table></div>
<!-- EDIT111 TABLE [40454-40736] -->
<p>
slides were measured with Serum(-) and Serum(+)<br/>
</p>
</div>
<!-- EDIT110 SECTION "2015.08.18" [39926-40788] -->
<h3 class="sectionedit112">2015.08.19</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocked spots 2x with 10 mg/mL BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> blocked slides with 10 mg/mL BSA for 30 min</div>
</li>
<li class="level1"><div class="li"> washed slides with PBS/diluted PB + 20 mM Imidazole/diluted PBS for 10 min </div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
Flush protocol Measuring with Serum (-) [not immunized] and Serum (+) [immunized]/a-GFP:
</p>
<div class="table sectionedit113"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> # </th><th class="col2"> Flowrate </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">780</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">780</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Serum (-)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4"> * </td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">Serum (+)/a-GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4"> * / 3ug/ml </td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">anti-Rabbit</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row10">
<td class="col0">Strep Cy5</td><td class="col1">10</td><td class="col2">30</td><td class="col3">6000</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row11">
<td class="col0">Buffer</td><td class="col1">11</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT113 TABLE [41120-41456] -->
</div>
<h4>Results</h4>
<div class="level4">
<p>
Serum (-) / Serum (+)
</p>
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/25/Freiburg_labjournal-roi_selection_20150819_315.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:roi_selection_20150819_315.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 315: Selection of ROIs</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/0/0f/Freiburg_labjournal-binding_curve_20150819_315.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:binding_curve_20150819_315.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 315: Binding curves</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/d/d1/Freiburg_labjournal-roi_selection_20150819_502.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:roi_selection_20150819_502.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 502: Selection of ROIs</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/6b/Freiburg_labjournal-binding_curve_20150819_502.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:binding_curve_20150819_502.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 502: Binding curves</div></div></div>
</div>
<!-- EDIT112 SECTION "2015.08.19" [40789-41845] -->
<h2 class="sectionedit114">Experiment 56: DNA on PDMS 4.0</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/PDMS_PDITC" title="protocols:pdms_pditc">PDMS-PDITC</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Cellfex_PDMS" title="protocols:cellfree_expression_on_pdms">DNA + Cellfree Expression on PDMS</a></div>
</li>
</ul>
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 PDMS slides for spotting with DNA</div>
</li>
</ul>
</div>
<!-- EDIT114 SECTION "Experiment 56: DNA on PDMS 4.0" [41846-42145] -->
<h3 class="sectionedit115">2015.08.17</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> used 2 flow cells made on 16.08.2015 with same pattern as flow cells used in iRIf (conic, one hole, one with pillars/one bad)</div>
</li>
<li class="level1"><div class="li"> washed PDMS slides with Ethanol and H<sub>2</sub>O</div>
</li>
<li class="level1"><div class="li"> activated slides in plasma generator with 30 L/h for 1 min</div>
</li>
<li class="level1"><div class="li"> incubated slides in APTES o/n at room temperature</div>
</li>
</ul>
</div>
<!-- EDIT115 SECTION "2015.08.17" [42146-42495] -->
<h3 class="sectionedit116">2015.08.18</h3>
<div class="level3">
<ul>
<li class="level1"><div class="li"> each slide was washed with ethanol</div>
</li>
<li class="level1"><div class="li"> a 5 min washing step in Ethanol followed</div>
</li>
<li class="level1"><div class="li"> after drying with wafergun slides were baked in oven (better one) for 90 min at 70°C</div>
</li>
<li class="level1"><div class="li"> slides were cooled down with wafergun</div>
</li>
<li class="level1"><div class="li"> incubation in PDITC for 2h at RT</div>
</li>
<li class="level1"><div class="li"> filled slideholder with EtOH and discarded it immediately</div>
</li>
<li class="level1"><div class="li"> washed 2x 5 min with EtOH in slideholder</div>
</li>
<li class="level1"><div class="li"> filled slideholder again with EtOH and discarded it immediately</div>
</li>
<li class="level1"><div class="li"> dried with wafergun</div>
</li>
<li class="level1"><div class="li"> flow cells were spotted with 1 µl spots and incubated over night at room temperature in humid atmosphere</div>
</li>
</ul>
<p>
spotting pattern:
<a class="media" href="https://static.igem.org/mediawiki/2015/0/00/Freiburg_labjournal-surchem-spotting_pattern_56.png" title="labjournal:surchem:spotting_pattern_56.png"><img alt="" class="media" src="https://static.igem.org/mediawiki/2015/0/00/Freiburg_labjournal-surchem-spotting_pattern_56.png" width="200"/></a>
</p>
<div class="table sectionedit117"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1</td><td class="col1">Cy5-HA-GFP-His-His</td><td class="col2">12.5 ng/µl</td>
</tr>
<tr class="row2">
<td class="col0">2</td><td class="col1">Cy5-HA-GFP-His-His</td><td class="col2">17.9 ng/µl</td>
</tr>
<tr class="row3">
<td class="col0">3</td><td class="col1">His-tYFP-Spy (lina104)</td><td class="col2">25 ng/µl</td>
</tr>
<tr class="row4">
<td class="col0">4</td><td class="col1">HA-GFP-Halo-His-His (Tobi)</td><td class="col2">25 ng/µl</td>
</tr>
<tr class="row5">
<td class="col0">5</td><td class="col1">Cy5-HA-GFP-His-His</td><td class="col2">22.4 ng/µl</td>
</tr>
<tr class="row6">
<td class="col0">6</td><td class="col1">PCR-Mix</td><td class="col2">-</td>
</tr>
</table></div>
<!-- EDIT117 TABLE [43152-43377] -->
</div>
<!-- EDIT116 SECTION "2015.08.18" [42496-43378] -->
<h3 class="sectionedit118">2015.08.19</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> expression was effected with 15 µl of Koko mix for 2h at 37°C</div>
</li>
<li class="level1"><div class="li"> flow cells were then checked for fluorescence with microscope of AG Eimer</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> for both flow cells no fluorescence could be observed </div>
</li>
</ul>
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/a/a5/Freiburg_labjournal-surchem-igem_150819_56a.png" title="labjournal:surchem:igem_150819_56a.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/a/a5/Freiburg_labjournal-surchem-igem_150819_56a.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/a/a5/Freiburg_labjournal-surchem-igem_150819_56a.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>56a</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/c/c7/Freiburg_labjournal-surchem-igem_150819_56b.png" title="labjournal:surchem:igem_150819_56b.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/c/c7/Freiburg_labjournal-surchem-igem_150819_56b.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/c/c7/Freiburg_labjournal-surchem-igem_150819_56b.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>56b</div></div></div>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> flow cells were disassembled and washed with PBS and H<sub>2</sub>O</div>
</li>
<li class="level1"><div class="li"> dried with wafergun</div>
</li>
<li class="level1"><div class="li"> expression was repeated with 15 µl of EasyExpress-Mix (AG Roth) at 37°C in humid atmosphere</div>
</li>
<li class="level1"><div class="li"> fluorescence was measured after 1h and 2h with microscope of AG Eimer</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> a few fluorescent spots could be observed</div>
</li>
</ul>
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/0/09/Freiburg_labjournal-surchem-igem_150819_rothmix_56a_1h.png" title="labjournal:surchem:igem_150819_rothmix_56a_1h.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/0/09/Freiburg_labjournal-surchem-igem_150819_rothmix_56a_1h.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/0/09/Freiburg_labjournal-surchem-igem_150819_rothmix_56a_1h.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>56a after 1h</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/e/e9/Freiburg_labjournal-surchem-igem_150819_rothmix_56a_2h.png" title="labjournal:surchem:igem_150819_rothmix_56a_2h.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/e/e9/Freiburg_labjournal-surchem-igem_150819_rothmix_56a_2h.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/e/e9/Freiburg_labjournal-surchem-igem_150819_rothmix_56a_2h.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>56a after 2h</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/6/63/Freiburg_labjournal-surchem-igem_150819_rothmix_56b_1h.png" title="labjournal:surchem:igem_150819_rothmix_56b_1h.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/63/Freiburg_labjournal-surchem-igem_150819_rothmix_56b_1h.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/6/63/Freiburg_labjournal-surchem-igem_150819_rothmix_56b_1h.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>56b after 1h</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/9/99/Freiburg_labjournal-surchem-igem_150819_rothmix_56b_2h.png" title="labjournal:surchem:igem_150819_rothmix_56b_2h.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/9/99/Freiburg_labjournal-surchem-igem_150819_rothmix_56b_2h.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/9/99/Freiburg_labjournal-surchem-igem_150819_rothmix_56b_2h.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>56b after 2h</div></div></div>
</div>
<!-- EDIT118 SECTION "2015.08.19" [43379-44417] -->
<h2 class="sectionedit119">Experiment 55: DNA on PDMS 3.0</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/PDMS_PDITC" title="protocols:pdms_pditc">PDMS-PDITC</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Cellfex_PDMS" title="protocols:cellfree_expression_on_pdms">DNA + Cellfree Expression on PDMS</a></div>
</li>
</ul>
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 PDMS slides for spotting with DNA</div>
</li>
</ul>
</div>
<!-- EDIT119 SECTION "Experiment 55: DNA on PDMS 3.0" [44418-44717] -->
<h3 class="sectionedit120">2015.08.15</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> used 2 flow cells (one with same pattern as flow cells used in iRIf and pillars, one with cavities and three lines)</div>
</li>
<li class="level1"><div class="li"> washed PDMS slides with Ethanol and H<sub>2</sub>O</div>
</li>
<li class="level1"><div class="li"> used correct APTES solution (1 mL APTES, 1 mL H<sub>2</sub>O in 18 mL Ethanol)</div>
</li>
<li class="level1"><div class="li"> activated slides in  Plasma Generator with 40 L/h for 1 min</div>
</li>
<li class="level1"><div class="li"> incubated slides in APTES o/n</div>
</li>
</ul>
</div>
<!-- EDIT120 SECTION "2015.08.15" [44718-45121] -->
<h3 class="sectionedit121">2015.08.16</h3>
<div class="level3">
<ul>
<li class="level1"><div class="li"> each slide was washed with ethanol</div>
</li>
<li class="level1"><div class="li"> a 5 min washing step in Ethanol followed</div>
</li>
<li class="level1"><div class="li"> after drying with wafergun slides were baked in oven (better one) for 90 min at 70°C</div>
</li>
<li class="level1"><div class="li"> slides were cooled down with wafergun</div>
</li>
<li class="level1"><div class="li"> incubation in PDITC for 3h at RT</div>
</li>
<li class="level1"><div class="li"> filled slideholder with EtOH and discarded it immediately</div>
</li>
<li class="level1"><div class="li"> washed 2x 5 min with EtOH in slideholder</div>
</li>
<li class="level1"><div class="li"> filled slideholder again with EtOH and discarded it immediately</div>
</li>
<li class="level1"><div class="li"> dried with wafergun</div>
</li>
<li class="level1"><div class="li"> cells were spotted with 3 µl spots and incubated over night at room temperature in humid atmosphere</div>
</li>
<li class="level1"><div class="li"> sample 1 was diluted 1:2 (original oncentration 44,8 ng/µl) in NaPi buffer 150 mM</div>
</li>
</ul>
<p>
spotting pattern:
</p>
<div class="thumb2 tleft" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/5/56/Freiburg_labjournal-surchem-spotting_pattern_55a.png" title="labjournal:surchem:spotting_pattern_55a.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/5/56/Freiburg_labjournal-surchem-spotting_pattern_55a.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/5/56/Freiburg_labjournal-surchem-spotting_pattern_55a.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>flow cell 55a</div></div></div><div class="thumb2 tleft" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/0/0e/Freiburg_labjournal-surchem-spotting_pattern_55b.png" title="labjournal:surchem:spotting_pattern_55b.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/0/0e/Freiburg_labjournal-surchem-spotting_pattern_55b.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/0/0e/Freiburg_labjournal-surchem-spotting_pattern_55b.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>flow cell 55b</div></div></div>
<p>
<br/>
<br/>
<br/>
</p>
<div class="table sectionedit122"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1</td><td class="col1">Cy5-HA-GFP-His-His</td><td class="col2">22,4 ng/µl</td>
</tr>
<tr class="row2">
<td class="col0">2</td><td class="col1">Control:HA-GFP-Halo-His-His</td><td class="col2">25 ng/µl</td>
</tr>
<tr class="row3">
<td class="col0">3</td><td class="col1">PCR-Mix of 1</td><td class="col2">-</td>
</tr>
</table></div>
<!-- EDIT122 TABLE [45955-46079] -->
</div>
<!-- EDIT121 SECTION "2015.08.16" [45122-46082] -->
<h3 class="sectionedit123">2015.08.17</h3>
<div class="level3">
<ul>
<li class="level1"><div class="li"> Petri dish with flow cells was put in oven at 60°C for 30 min</div>
</li>
<li class="level1"><div class="li"> Flow cells were then washed with aqua dest. and dried with wafergun</div>
</li>
<li class="level1"><div class="li"> Expression was effected for 20, 40 and 130 min with AG Roth expression mix at 37°C (not sealed, no humid atmosphere)</div>
</li>
<li class="level1"><div class="li"> flow cells were checked for fluorescence in microscope of AG Eimer</div>
</li>
<li class="level1"><div class="li"> afterwards flow cells were washed with 2 ml PBS and aqua dest. and thene dried with wafergun</div>
</li>
<li class="level1"><div class="li"> stored in petri dish sealed with parafilm at 4°C</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> Expression of our GFP conctruct and the control construct could be observed on both flow cells</div>
</li>
<li class="level1"><div class="li"> Expressed GFP seems to diffuse with the time. This might be due to the transport of the slides from the incubator to the microscope and back.</div>
</li>
</ul>
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/2/20/Freiburg_labjournal-surchem-20150817_exp55a_20min.png" title="labjournal:surchem:20150817_exp55a_20min.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/20/Freiburg_labjournal-surchem-20150817_exp55a_20min.png" width="300"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/2/20/Freiburg_labjournal-surchem-20150817_exp55a_20min.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>55a after 20 min incubation time</div></div></div><div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/f/f3/Freiburg_labjournal-surchem-20150817_exp55a_40min.png" title="labjournal:surchem:20150817_exp55a_40min.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/f3/Freiburg_labjournal-surchem-20150817_exp55a_40min.png" width="300"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/f/f3/Freiburg_labjournal-surchem-20150817_exp55a_40min.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>55a after 40 min incubation time</div></div></div><div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/9/9a/Freiburg_labjournal-surchem-20150817_exp55a_130min.png" title="labjournal:surchem:20150817_exp55a_130min.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/9/9a/Freiburg_labjournal-surchem-20150817_exp55a_130min.png" width="300"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/9/9a/Freiburg_labjournal-surchem-20150817_exp55a_130min.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>55a after 130 min incubation time</div></div></div><div class="thumb2 trien" style="width:310px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/4/46/Freiburg_labjournal-surchem-20150817_exp55b.png" title="labjournal:surchem:20150817_exp55b.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/46/Freiburg_labjournal-surchem-20150817_exp55b.png" width="300"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/4/46/Freiburg_labjournal-surchem-20150817_exp55b.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>55b after 20 min incubation time</div></div></div>
</div>
<!-- EDIT123 SECTION "2015.08.17" [46083-47206] -->
<h3 class="sectionedit124">2015.08.18</h3>
<div class="level3">
<ul>
<li class="level1"><div class="li"> Expression was effected for 2h with Koko expression mix at 37°C (sealed, humid atmosphere)</div>
</li>
<li class="level1"><div class="li"> flow cells were checked for fluorescence in microscope of AG Eimer</div>
</li>
<li class="level1"><div class="li"> afterwards flow cells were washed with 2 ml PBS and aqua dest. and thene dried with wafergun</div>
</li>
<li class="level1"><div class="li"> stored in petri dish sealed with parafilm at 4°C</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> no fluorescence could be observed for both flow  cells as well as for Tobis 4 flow cells</div>
</li>
</ul>
<div class="thumb2 trien" style="width:310px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/6/68/Freiburg_labjournal-surchem-20150818_exp55a_2h.png" title="labjournal:surchem:20150818_exp55a_2h.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/68/Freiburg_labjournal-surchem-20150818_exp55a_2h.png" width="300"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/6/68/Freiburg_labjournal-surchem-20150818_exp55a_2h.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>55a after 2h</div></div></div><div class="thumb2 trien" style="width:310px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/b/bd/Freiburg_labjournal-surchem-20150818_exp55b_2h.png" title="labjournal:surchem:20150818_exp55b_2h.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/b/bd/Freiburg_labjournal-surchem-20150818_exp55b_2h.png" width="300"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/b/bd/Freiburg_labjournal-surchem-20150818_exp55b_2h.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>55b after 2h</div></div></div>
</div>
<!-- EDIT124 SECTION "2015.08.18" [47207-47789] -->
<h2 class="sectionedit125">Experiment 54b: Ni-NTA iRIf slides - rabbit Serum</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT125 SECTION "Experiment 54b: Ni-NTA iRIf slides - rabbit Serum" [47790-47974] -->
<h3 class="sectionedit126">2015.08.15</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 4 PDITC iRIf slides </div>
</li>
<li class="level1"><div class="li"> slides <a class="bad-entry" href="" id="" title="FIXME: Nothing to link to!  First create a default subjectindex table for entries in this index section (e.g. {{subjectindex&gt;...section=?;default}}  See plugin help file online for details)">:</a>
 504, 500, 216, 208</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 4 PDITC iRIf slides were prepared</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
</ul>
</div>
<!-- EDIT126 SECTION "2015.08.15" [47975-48241] -->
<h3 class="sectionedit127">2015.08.17</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slides were sandwiched with 60 µL NTA-solution and incubated o/n at 4 °C</div>
</li>
</ul>
</div>
<!-- EDIT127 SECTION "2015.08.17" [48242-48366] -->
<h3 class="sectionedit128">2015.08.18</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slides were blocked in APTES blocking solution for 1 h</div>
</li>
<li class="level1"><div class="li"> slides were incubated in NiSO&lt;Sub&gt;4&lt;/Sub&gt; for 1 h</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS/diluted PBS</div>
</li>
<li class="level1"><div class="li"> 3 µL of cell free expressed GFP, GFP Lysate, GFP desalt, bBSA, BSA and mCherry were spotted.</div>
</li>
</ul>
<p>
spotting pattern:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<div class="table sectionedit129"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1-2 </td><td class="col1">His-GFP (cellfree expressed))</td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0"> 3 </td><td class="col1"> His-GFP desalt. </td><td class="col2"> 0.2 mg/ml </td>
</tr>
<tr class="row3">
<td class="col0"> 4-5 </td><td class="col1">neg. control (cellfree Expression without DNA)</td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0"> 6-7 </td><td class="col1"> His-mCherry lysate </td><td class="col2"> 1:10 </td>
</tr>
<tr class="row5">
<td class="col0"> 8-9 </td><td class="col1"> His-GFP lysate </td><td class="col2"> 1:10 </td>
</tr>
<tr class="row6">
<td class="col0"> 10 </td><td class="col1">BSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
<tr class="row7">
<td class="col0"> 11 </td><td class="col1">bBSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
</table></div>
<!-- EDIT129 TABLE [48750-49032] -->
<p>
3 of the slides (208, 216, 504) were measured with Serum(-) and Serum(+)<br/>

1 of the slides (500) was measured with Serum(-) and anti-GFP<br/>

(slide from Exp. 50c was spotted similar and also measured with Serum(-) and anti-GFP)<br/>
</p>
</div>
<!-- EDIT128 SECTION "2015.08.18" [48367-49262] -->
<h3 class="sectionedit130">2015.08.19</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocked spots 2x with 10 mg/mL BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> blocked slides with 10 mg/mL BSA for 30 min</div>
</li>
<li class="level1"><div class="li"> washed slides with PBS/diluted PB + 20 mM Imidazole/diluted PBS for 10 min </div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
Flush protocol Measuring with Serum (-) [not immunized] and Serum (+) [immunized]/a-GFP:
</p>
<div class="table sectionedit131"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> # </th><th class="col2"> Flowrate </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">780</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">780</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Serum (-)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4"> * </td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">Serum (+)/a-GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4"> * / 3ug/ml </td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">anti-Rabbit</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row10">
<td class="col0">Strep Cy5</td><td class="col1">10</td><td class="col2">30</td><td class="col3">6000</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row11">
<td class="col0">Buffer</td><td class="col1">11</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT131 TABLE [49594-49930] -->
</div>
<h4>Results</h4>
<div class="level4">
<p>
Serum (-) / Serum (+)
</p>
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/c/cc/Freiburg_labjournal-roi_selection_20150819_208.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:roi_selection_20150819_208.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 208: Selection of ROIs</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/9/92/Freiburg_labjournal-binding_curve_20150819_208.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:binding_curve_20150819_208.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 208: Binding curves</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/8/8d/Freiburg_labjournal-roi_selection_20150819_504.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:roi_selection_20150819_504.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 502: Selection of ROIs</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/6e/Freiburg_labjournal-binding_curve_20150819_504.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:binding_curve_20150819_504.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 502: Binding curves</div></div></div><hr/>
<p>
Serum (-) / a-GFP
</p>
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/9/9e/Freiburg_labjournal-roi_selection_20150819_424.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:roi_selection_20150819_424.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 424: Selection of ROIs</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/d/d8/Freiburg_labjournal-binding_curve_20150819_424.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:binding_curve_20150819_424.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 424: Binding curves</div></div></div><hr/>
<p>
The relative intensity shift for the three approaches (Serum(-)Serum(+) on Ni-NTA ; Serum(-)Serum(+) on PDITC; Serum(-)a-GFP on Ni-NTA)
were compared. As expected and seen in the binding curves cellfree expression Mix (His-GFP) shows no signal on PDITC (other proteins within the
mix bind to the surface) but on specific Ni-NTA surface. Taken amount of serum performs less (2-3 fold) than 3 ug/ml a-GFP.
</p>
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/d/d9/Freiburg_labjournal-evaluation_rel._intshift_rabbitserum.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:evaluation_rel._intshift_rabbitserum.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Evaluation of the relative intensity shift. From left to right spot 1-11. For each spot rel. intensity shift is plotted with bars
for Serum(-)flush and Serum(+)/a-GFP</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/0/02/Freiburg_labjournal-mcherry_emission_rabbitserum.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:mcherry_emission_rabbitserum.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Emission within 532nm channel (mCherry) after iRIf measurement. Averaged over all samples.</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/9/96/Freiburg_labjournal-cy5_emission_rabbitserum.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:cy5_emission_rabbitserum.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Emission within 635nm channel (Cy5) after iRIf measurement. Averaged over all samples</div></div></div>
</div>
<!-- EDIT130 SECTION "2015.08.19" [49263-51460] -->
<h2 class="sectionedit132">Experiment 54a: PDITC iRIf slides - own device</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT132 SECTION "Experiment 54a: PDITC iRIf slides - own device" [51461-51642] -->
<h3 class="sectionedit133">2015.08.15</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 1 PDITC iRIf slides </div>
</li>
<li class="level1"><div class="li"> slides <a class="bad-entry" href="" id="" title="FIXME: Nothing to link to!  First create a default subjectindex table for entries in this index section (e.g. {{subjectindex&gt;...section=?;default}}  See plugin help file online for details)">:</a>
 467</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 1 PDITC iRIf slides were prepared</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
</ul>
</div>
<!-- EDIT133 SECTION "2015.08.15" [51643-51894] -->
<h3 class="sectionedit134">2015.08.17</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 1 slide was spotted alternating with bBSA and BSA for measurement in own device, in total 14 spots</div>
</li>
</ul>
</div>
<!-- EDIT134 SECTION "2015.08.17" [51895-52045] -->
<h3 class="sectionedit135">2015.08.18</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slide for own device was blocked (2x 10 mg/mL BSA on spots for 5 min, 1x in 10 mg/mL BSA for 30 min)</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
</div>
<!-- EDIT135 SECTION "2015.08.18" [52046-52213] -->
<h2 class="sectionedit136">Experiment 53b: Plasma activated slides with PLL-PEG-HTL from Piehler</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Halo_Piehler" title="protocols:halosurface_piehler">Halosurface from AG Piehler</a></div>
</li>
</ul>
</div>
<!-- EDIT136 SECTION "Experiment 53b: Plasma activated slides with PLL-PEG-HTL from Piehler" [52214-52429] -->
<h3 class="sectionedit137">2015.08.14</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> second try with halo surface</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> activated slides <strong>253</strong> and <strong>254</strong> with 40 L/h for 5 min</div>
</li>
<li class="level1"><div class="li"> cross-sandwiched slides with 15 µL PLL-PEG-HTL</div>
</li>
<li class="level1"><div class="li"> incubated slides for 10 min</div>
</li>
<li class="level1"><div class="li"> washed slides with Aqua dest and stored them in slideholder o/n (no N<sub>2</sub>-atmosphere)</div>
</li>
</ul>
</div>
<!-- EDIT137 SECTION "2015.08.14" [52430-52778] -->
<h3 class="sectionedit138">2015.08.16</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocked slides in 10 mg/ml BSA for 1h</div>
</li>
<li class="level1"><div class="li"> washed slides with aqua dest. and dried with nitrogengun</div>
</li>
<li class="level1"><div class="li"> spotted slides and incubated for 1 h at roomtemperature</div>
</li>
</ul>
<p>
Spotting:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<div class="table sectionedit139"><table class="inline">
<tr class="row0">
<th class="col0"> Spot </th><th class="col1"> Protein </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-2</td><td class="col1"> Halo GFP</td><td class="col2"> 0.5 mg/ml</td>
</tr>
<tr class="row2">
<td class="col0">3-4</td><td class="col1"> Halo GFP </td><td class="col2">0.1 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">5-6</td><td class="col1"> Halo GFP</td><td class="col2">0.02 mg/ml</td>
</tr>
<tr class="row4">
<td class="col0">7</td><td class="col1"> bBSA</td><td class="col2">0.2 mg/ml</td>
</tr>
<tr class="row5">
<td class="col0">8</td><td class="col1"> Halo mCherry </td><td class="col2">0.5 mg/ml</td>
</tr>
<tr class="row6">
<td class="col0">9</td><td class="col1"> His-GFP (Max)</td><td class="col2">0.5 mg/ml</td>
</tr>
<tr class="row7">
<td class="col0">10</td><td class="col1"> His Halo GFP (Piehler, pos. control) </td><td class="col2">0.25 mg/ml</td>
</tr>
<tr class="row8">
<td class="col0">11</td><td class="col1"> His GFP (Piehler, neg. control)</td><td class="col2">0.25 mg/ml</td>
</tr>
</table></div>
<!-- EDIT139 TABLE [53064-53361] --><ul>
<li class="level1"><div class="li"> blocked wells twice with 10 mg/ml BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> blocked slides 30 min in 10 mg/ml BSA</div>
</li>
<li class="level1"><div class="li"> washed slides with water and dried with nitrogengun</div>
</li>
<li class="level1"><div class="li"> measured fluorescence intensity with fluorescence microscope</div>
</li>
<li class="level1"><div class="li"> measurement in iRIf</div>
</li>
</ul>
<div class="table sectionedit140"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer (PBS)</td><td class="col1">1</td><td class="col2">60</td><td class="col3">720</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0 leftalign">BSA	</td><td class="col1">2</td><td class="col2">60</td><td class="col3">720</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer	(PBS)</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Anti GFP (biotinylated, goat)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer	(PBS)</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">StrepCy5</td><td class="col1">6</td><td class="col2">60</td><td class="col3">300</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer	(PBS)</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT140 TABLE [53603-53883] -->
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/e/ee/Freiburg_labjournal-surchem-exp_53b_slide254.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_53b_slide254.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 254: Quotion picture</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/27/Freiburg_labjournal-surchem-exp_53b_slide254_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_53b_slide254_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 254: Picture obtained from fluorescence scanner with enhanced brightness. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><hr/>
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/45/Freiburg_labjournal-surchem-exp_53b_slide253.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_53b_slide253.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 253: Quotion picture</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/0/03/Freiburg_labjournal-surchem-exp_53b_slide253_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_53b_slide253_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 253: Picture obtained from fluorescence scanner with enhanced brightness. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div>
</div>
<!-- EDIT138 SECTION "2015.08.16" [52779-54474] -->
<h2 class="sectionedit141">Experiment 53a: Plasma activated slides with abcr-silane</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Halo_ABCR" title="protocols:halosurface_abcr">Halosurface from ABCR</a></div>
</li>
</ul>
</div>
<!-- EDIT141 SECTION "Experiment 53a: Plasma activated slides with abcr-silane" [54475-54668] -->
<h3 class="sectionedit142">2015.08.14</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> second try with halo surface</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> activated slides <strong>012</strong> and <strong>215</strong> with 40 L/h for 5 min</div>
</li>
<li class="level1"><div class="li"> sandwiched slides with 80 µL abcr-silane</div>
</li>
<li class="level1"><div class="li"> incubated slides for 1 h</div>
</li>
<li class="level1"><div class="li"> washed slides with Aqua dest and stored them in slideholder o/n (no N<sub>2</sub>-atmosphere)</div>
</li>
</ul>
</div>
<!-- EDIT142 SECTION "2015.08.14" [54669-55008] -->
<h3 class="sectionedit143">2015.08.16</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocked slides in 10 mg/ml BSA for 1h</div>
</li>
<li class="level1"><div class="li"> washed slides with aqua dest. and dried with nitrogengun</div>
</li>
<li class="level1"><div class="li"> spotted slides and incubated for 1 h at roomtemperature</div>
</li>
</ul>
<p>
Spotting:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<div class="table sectionedit144"><table class="inline">
<tr class="row0">
<th class="col0"> Spot </th><th class="col1"> Protein </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-2</td><td class="col1"> Halo GFP</td><td class="col2"> 0.5 mg/ml</td>
</tr>
<tr class="row2">
<td class="col0">3-4</td><td class="col1"> Halo GFP </td><td class="col2">0.1 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">5-6</td><td class="col1"> Halo GFP</td><td class="col2">0.02 mg/ml</td>
</tr>
<tr class="row4">
<td class="col0">7</td><td class="col1"> bBSA</td><td class="col2">0.2 mg/ml</td>
</tr>
<tr class="row5">
<td class="col0">8</td><td class="col1"> Halo mCherry </td><td class="col2">0.5 mg/ml</td>
</tr>
<tr class="row6">
<td class="col0">9</td><td class="col1"> His-GFP (Max)</td><td class="col2">0.5 mg/ml</td>
</tr>
<tr class="row7">
<td class="col0">10</td><td class="col1"> His Halo GFP (Piehler, pos. control) </td><td class="col2">0.25 mg/ml</td>
</tr>
<tr class="row8">
<td class="col0">11</td><td class="col1"> His GFP (Piehler, neg. control)</td><td class="col2">0.25 mg/ml</td>
</tr>
</table></div>
<!-- EDIT144 TABLE [55294-55591] --><ul>
<li class="level1"><div class="li"> blocked wells twice with 10 mg/ml BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> blocked slides 30 min in 10 mg/ml BSA</div>
</li>
<li class="level1"><div class="li"> washed slides with water and dried with nitrogengun</div>
</li>
<li class="level1"><div class="li"> measured fluorescence intensity with fluorescence microscope</div>
</li>
<li class="level1"><div class="li"> measurement in iRIf</div>
</li>
</ul>
<div class="table sectionedit145"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer (PBS)</td><td class="col1">1</td><td class="col2">60</td><td class="col3">720</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0 leftalign">BSA	</td><td class="col1">2</td><td class="col2">60</td><td class="col3">720</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer	(PBS)</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Anti GFP (biotinylated, goat)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer	(PBS)</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">StrepCy5</td><td class="col1">6</td><td class="col2">60</td><td class="col3">300</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer	(PBS)</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT145 TABLE [55833-56113] -->
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/f1/Freiburg_labjournal-surchem-exp_53a_slide012.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_53a_slide012.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 012: Quotion picture</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/7e/Freiburg_labjournal-surchem-exp_53a_slide012_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_53a_slide012_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 012: Picture obtained from fluorescence scanner with enhanced brightness. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><hr/>
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/6a/Freiburg_labjournal-surchem-exp_53a_slide215.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_53a_slide215.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 215: Quotion picture</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/2f/Freiburg_labjournal-surchem-exp_53a_slide215_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_53a_slide215_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 215: Picture obtained from fluorescence scanner with enhanced brightness. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div>
</div>
<!-- EDIT143 SECTION "2015.08.16" [55009-56706] -->
<h2 class="sectionedit146">Experiment 52c: PDITC Slides</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT146 SECTION "Experiment 52c: PDITC Slides" [56707-56869] -->
<h3 class="sectionedit147">2015.08.14</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> washed 1 iRIf slides</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 45 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
<li class="level1"><div class="li"> stored slides at 4 °C o/n</div>
</li>
</ul>
</div>
<!-- EDIT147 SECTION "2015.08.14" [56870-57088] -->
<h3 class="sectionedit148">2015.08.17</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slide was spotted alternating with bBSA and BSA for measurement in own device, in total 14 spots</div>
</li>
</ul>
</div>
<!-- EDIT148 SECTION "2015.08.17" [57089-57236] -->
<h3 class="sectionedit149">2015.08.18</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slide was blocked (2x 10 mg/mL BSA on spots for 5 min, 1x in 10 mg/mL BSA for 30 min)</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
</div>
<!-- EDIT149 SECTION "2015.08.18" [57237-57390] -->
<h2 class="sectionedit150">Experiment 52b: PDITC Slides with Wiesmüller Halo-Linker 2</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Halo_Wiesmueller" title="protocols:halosurface_wiesmueller">Halosurface from Wiesmüller</a></div>
</li>
</ul>
</div>
<!-- EDIT150 SECTION "Experiment 52b: PDITC Slides with Wiesmüller Halo-Linker 2" [57391-57652] -->
<h3 class="sectionedit151">2015.08.14</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> washed 2 iRIf slides</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 45 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
<li class="level1"><div class="li"> stored slides at 4 °C o/n</div>
</li>
</ul>
<ul>
<li class="level1"><div class="li"> slide <strong>423</strong> and <strong>287</strong> were incubated with Wiesmüller Halo-Linker 2 o/n in slideholder</div>
</li>
</ul>
</div>
<!-- EDIT151 SECTION "2015.08.14" [57653-57967] -->
<h3 class="sectionedit152">2015.08.16</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> washed slides with Aqua dest. and dried with nitrogengun</div>
</li>
<li class="level1"><div class="li"> blocked slides in 10 mg/ml BSA for 1h</div>
</li>
<li class="level1"><div class="li"> washed slides with aqua dest. and dried with nitrogengun</div>
</li>
<li class="level1"><div class="li"> spotted slides and incubated for 1 h at roomtemperature</div>
</li>
</ul>
<p>
Spotting:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<div class="table sectionedit153"><table class="inline">
<tr class="row0">
<th class="col0"> Spot </th><th class="col1"> Protein </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-2</td><td class="col1"> Halo GFP</td><td class="col2"> 0.5 mg/ml</td>
</tr>
<tr class="row2">
<td class="col0">3-4</td><td class="col1"> Halo GFP </td><td class="col2">0.1 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">5-6</td><td class="col1"> Halo GFP</td><td class="col2">0.02 mg/ml</td>
</tr>
<tr class="row4">
<td class="col0">7</td><td class="col1"> bBSA</td><td class="col2">0.2 mg/ml</td>
</tr>
<tr class="row5">
<td class="col0">8</td><td class="col1"> Halo mCherry </td><td class="col2">0.5 mg/ml</td>
</tr>
<tr class="row6">
<td class="col0">9</td><td class="col1"> His-GFP (Max)</td><td class="col2">0.5 mg/ml</td>
</tr>
<tr class="row7">
<td class="col0">10</td><td class="col1"> His Halo GFP (Piehler, pos. control) </td><td class="col2">0.25 mg/ml</td>
</tr>
<tr class="row8">
<td class="col0">11</td><td class="col1"> His GFP (Piehler, neg. control)</td><td class="col2">0.25 mg/ml</td>
</tr>
</table></div>
<!-- EDIT153 TABLE [58314-58611] --><ul>
<li class="level1"><div class="li"> blocked wells twice with 10 mg/ml BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> blocked slides 30 min in 10 mg/ml BSA</div>
</li>
<li class="level1"><div class="li"> washed slides with water and dried with nitrogengun</div>
</li>
<li class="level1"><div class="li"> measured fluorescence intensity with fluorescence microscope</div>
</li>
<li class="level1"><div class="li"> measurement in iRIf</div>
</li>
</ul>
<div class="table sectionedit154"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer (PBS)</td><td class="col1">1</td><td class="col2">60</td><td class="col3">720</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0 leftalign">BSA	</td><td class="col1">2</td><td class="col2">60</td><td class="col3">720</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer	(PBS)</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Anti GFP (biotinylated, goat)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer	(PBS)</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">StrepCy5</td><td class="col1">6</td><td class="col2">60</td><td class="col3">300</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer	(PBS)</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT154 TABLE [58853-59133] -->
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/1/1b/Freiburg_labjournal-surchem-exp_52b_slide423.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_52b_slide423.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 423: Quotion picture</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/f1/Freiburg_labjournal-surchem-exp_52b_slide423_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_52b_slide423_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 423: Picture obtained from fluorescence scanner with enhanced brightness. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><hr/>
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/d/df/Freiburg_labjournal-surchem-exp_52b_slide287.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_52b_slide287.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 287: Quotion picture</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/3/32/Freiburg_labjournal-surchem-exp_52b_slide287_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_52b_slide287_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 287: Picture obtained from fluorescence scanner with enhanced brightness. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div>
</div>
<!-- EDIT152 SECTION "2015.08.16" [57968-59726] -->
<h2 class="sectionedit155">Experiment 52a: PDITC Slides with Wiesmüller Halo-Linker 1</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Halo_Wiesmueller" title="protocols:halosurface_wiesmueller">Halosurface from Wiesmüller</a></div>
</li>
</ul>
</div>
<!-- EDIT155 SECTION "Experiment 52a: PDITC Slides with Wiesmüller Halo-Linker 1" [59727-59988] -->
<h3 class="sectionedit156">2015.08.14</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> washed 2 iRIf slides</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 40 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 45 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
<li class="level1"><div class="li"> stored slides at 4 °C o/n</div>
</li>
</ul>
<ul>
<li class="level1"><div class="li"> slide <strong>426</strong> and <strong>466</strong> were incubated with Wiesmüller Halo-Linker 1 o/n in slideholder</div>
</li>
</ul>
</div>
<!-- EDIT156 SECTION "2015.08.14" [59989-60303] -->
<h3 class="sectionedit157">2015.08.16</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> washed slides with Aqua dest. and dried with nitrogengun</div>
</li>
<li class="level1"><div class="li"> blocked slides in 10 mg/ml BSA for 1h</div>
</li>
<li class="level1"><div class="li"> washed slides with aqua dest. and dried with nitrogengun</div>
</li>
<li class="level1"><div class="li"> spotted slides and incubated for 1 h at roomtemperature</div>
</li>
</ul>
<p>
Spotting:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<div class="table sectionedit158"><table class="inline">
<tr class="row0">
<th class="col0"> Spot </th><th class="col1"> Protein </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-2</td><td class="col1"> Halo GFP</td><td class="col2"> 0.5 mg/ml</td>
</tr>
<tr class="row2">
<td class="col0">3-4</td><td class="col1"> Halo GFP </td><td class="col2">0.1 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">5-6</td><td class="col1"> Halo GFP</td><td class="col2">0.02 mg/ml</td>
</tr>
<tr class="row4">
<td class="col0">7</td><td class="col1"> bBSA</td><td class="col2">0.2 mg/ml</td>
</tr>
<tr class="row5">
<td class="col0">8</td><td class="col1"> Halo mCherry </td><td class="col2">0.5 mg/ml</td>
</tr>
<tr class="row6">
<td class="col0">9</td><td class="col1"> His-GFP (Max)</td><td class="col2">0.5 mg/ml</td>
</tr>
<tr class="row7">
<td class="col0">10</td><td class="col1"> His Halo GFP (Piehler, pos. control) </td><td class="col2">0.25 mg/ml</td>
</tr>
<tr class="row8">
<td class="col0">11</td><td class="col1"> His GFP (Piehler, neg. control)</td><td class="col2">0.25 mg/ml</td>
</tr>
</table></div>
<!-- EDIT158 TABLE [60650-60947] --><ul>
<li class="level1"><div class="li"> blocked wells twice with 10 mg/ml BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> blocked slides 30 min in 10 mg/ml BSA</div>
</li>
<li class="level1"><div class="li"> washed slides with water and dried with nitrogengun</div>
</li>
<li class="level1"><div class="li"> measured fluorescence intensity with fluorescence microscope</div>
</li>
<li class="level1"><div class="li"> measurement in iRIf</div>
</li>
</ul>
<div class="table sectionedit159"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer (PBS)</td><td class="col1">1</td><td class="col2">60</td><td class="col3">720</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0 leftalign">BSA	</td><td class="col1">2</td><td class="col2">60</td><td class="col3">720</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer	(PBS)</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Anti GFP (biotinylated, goat)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer	(PBS)</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">StrepCy5</td><td class="col1">6</td><td class="col2">60</td><td class="col3">300</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer	(PBS)</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT159 TABLE [61189-61469] -->
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/c/c7/Freiburg_labjournal-surchem-exp_52a_slide426.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_52a_slide426.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 426: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/62/Freiburg_labjournal-surchem-exp_52a_slide426_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_52a_slide426_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 426: Picture obtained from fluorescence scanner with enhanced brightness. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/1/11/Freiburg_labjournal-surchem-exp_52a_slide426_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_52a_slide426_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 426: Binding curves</div></div></div><hr/>
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/0/01/Freiburg_labjournal-surchem-exp_52a_slide466.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_52a_slide466.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 466: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/40/Freiburg_labjournal-surchem-exp_52a_slide466_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_52a_slide466_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 466: Picture obtained from fluorescence scanner with enhanced brightness. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/41/Freiburg_labjournal-surchem-exp_52a_slide466_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_52a_slide466_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 466: Binding curves</div></div></div>
</div>
<!-- EDIT157 SECTION "2015.08.16" [60304-62247] -->
<h2 class="sectionedit160">Experiment 47b/49c: PDITC for iRIF/new Device</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT160 SECTION "Experiment 47b/49c: PDITC for iRIF/new Device" [62248-62428] -->
<h3 class="sectionedit161">2015.08.13</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> test if antibody binding is detectable with self built iRIf setup </div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> used two already prepared iRIf PDITC slides from Exp. 47b (265) and 49c (503)</div>
</li>
<li class="level1"><div class="li"> spotted bBSA (200 µg/ml) and BSA (10 mg/ml) in an alternating pattern on slide 503</div>
</li>
<li class="level1"><div class="li"> spotted anti HCV antibodys (rabbit, polyclonal, 500 µg/ml)and BSA (10 mg/ml) an alternating pattern on slide 265</div>
</li>
<li class="level1"><div class="li"> spotted 18 spots in total on each slide</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
</div>
<!-- EDIT161 SECTION "2015.08.13" [62429-62919] -->
<h2 class="sectionedit162">Experiment 51: DNA on PDMS 2.0</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used : <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/PDMS_PDITC" title="protocols:pdms_pditc">PDMS-PDITC</a></div>
</li>
</ul>
</div>
<!-- EDIT162 SECTION "Experiment 51: DNA on PDMS 2.0" [62920-63020] -->
<h3 class="sectionedit163">2015.08.12</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> repeat immobilization of Cy5-labeled DNA on PDMS slides with correct APTES solution</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> used 2 flow chamber slides with same pattern as flow Chambers used in iRIf (conic, one hole)</div>
</li>
<li class="level1"><div class="li"> washed PDMS slides with Ethanol and H<sub>2</sub>O</div>
</li>
<li class="level1"><div class="li"> used correct APTES solution (1 mL APTES, 1 mL H<sub>2</sub>O in 18 mL Ethanol)</div>
</li>
<li class="level1"><div class="li"> activated slides in  Plasma Generator with 40 L/h for 1 min</div>
</li>
<li class="level1"><div class="li"> incubated slides in APTES o/n</div>
</li>
</ul>
</div>
<!-- EDIT163 SECTION "2015.08.12" [63021-63513] -->
<h3 class="sectionedit164">2015.08.13</h3>
<div class="level3">
<ul>
<li class="level1"><div class="li"> each slide was washed with ethanol</div>
</li>
<li class="level1"><div class="li"> a 5 min washing step in Ethanol followed</div>
</li>
<li class="level1"><div class="li"> after drying with wafergun slides were baked in oven for 90 min at 70°C</div>
</li>
<li class="level1"><div class="li"> slides were cooled down with wafergun</div>
</li>
<li class="level1"><div class="li"> incubation in PDITC for 5h40min at RT</div>
</li>
<li class="level1"><div class="li"> filled slideholder with EtOH and discarded it immediately</div>
</li>
<li class="level1"><div class="li"> washed 2x 5 min with EtOH in slideholder</div>
</li>
<li class="level1"><div class="li"> filled slideholder again with EtOH and discarded it immediately</div>
</li>
<li class="level1"><div class="li"> dried with wafergun</div>
</li>
<li class="level1"><div class="li"> 1 slide (51a) was spotted with 1 µl spots and incubated over night</div>
</li>
<li class="level1"><div class="li"> sample 1 was diluted 1:4,64 (original oncentration 116 ng/µl) in NaPi buffer 150 mM</div>
</li>
</ul>
<p>
spotting pattern 51a:
<a class="media" href="https://static.igem.org/mediawiki/2015/5/5b/Freiburg_labjournal-spotting_pattern_pdms_1.png" title="labjournal:spotting_pattern_pdms_1.png"><img align="left" alt="" class="medialeft" src="https://static.igem.org/mediawiki/2015/5/5b/Freiburg_labjournal-spotting_pattern_pdms_1.png" width="200"/></a>
</p>
<div class="table sectionedit165"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1</td><td class="col1">His-tYFP-Spy (lina104) (top 0.5 µl, bottom spot 1 µl)</td><td class="col2">25 ng/µl</td>
</tr>
<tr class="row2">
<td class="col0">2</td><td class="col1">His-tYFP-Halo (lina105)</td><td class="col2">20 ng/µl</td>
</tr>
<tr class="row3">
<td class="col0">3</td><td class="col1">HA-GFP-His<sub>6</sub>-His<sub>6</sub></td><td class="col2">28.7 ng/µl</td>
</tr>
<tr class="row4">
<td class="col0">4</td><td class="col1">Control by Tobi: HA-GFP-Halo-His</td><td class="col2">25 ng/µl</td>
</tr>
</table></div>
<!-- EDIT165 TABLE [64214-64452] --><ul>
<li class="level1"><div class="li"> slide (b) was spotted with 3 µl spots and incubated for 2 nights at room temperature</div>
</li>
<li class="level1"><div class="li"> sample 1 was diluted 1:4,64 (original concentration 116 ng/µl) in NaPi buffer 150 mM</div>
</li>
</ul>
<p>
spotting pattern 51b:
<a class="media" href="https://static.igem.org/mediawiki/2015/5/5b/Freiburg_labjournal-spotting_pattern_pdms_1.png" title="labjournal:spotting_pattern_pdms_1.png"><img align="left" alt="" class="medialeft" src="https://static.igem.org/mediawiki/2015/5/5b/Freiburg_labjournal-spotting_pattern_pdms_1.png" width="200"/></a>
</p>
<div class="table sectionedit166"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1</td><td class="col1">His-tYFP-Spy (lina104)</td><td class="col2">25 ng/µl</td>
</tr>
<tr class="row2">
<td class="col0">2</td><td class="col1">His-tYFP-Halo (lina105)</td><td class="col2">20 ng/µl</td>
</tr>
<tr class="row3">
<td class="col0">3</td><td class="col1">HA-GFP-His<sub>6</sub>-His<sub>6</sub></td><td class="col2">28.7 ng/µl</td>
</tr>
<tr class="row4">
<td class="col0">4</td><td class="col1">Control by Tobi: HA-GFP-Halo-HisHis</td><td class="col2">25 ng/µl</td>
</tr>
</table></div>
<!-- EDIT166 TABLE [64708-64916] -->
</div>
<!-- EDIT164 SECTION "2015.08.13" [63514-64917] -->
<h3 class="sectionedit167">2015.08.14</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> DNA was let dry in</div>
</li>
<li class="level1"><div class="li"> slide 51a was washed with H<sub>2</sub>O and dried with wafergun</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slide 51a was measured in Microarray Scanner for Cy5</div>
</li>
<li class="level1"><div class="li"> no spots could be seen  </div>
</li>
</ul>
<p>
<a class="media" href="https://static.igem.org/mediawiki/2015/f/ff/Freiburg_labjournal-surchem-20150814_exp51a_532.png" title="labjournal:surchem:20150814_exp51a_532.png"><img align="left" alt="532 nm" class="medialeft" src="https://static.igem.org/mediawiki/2015/f/ff/Freiburg_labjournal-surchem-20150814_exp51a_532.png" title="532 nm" width="100"/></a>
<a class="media" href="https://static.igem.org/mediawiki/2015/5/5b/Freiburg_labjournal-surchem-20150814_exp51a_635.png" title="labjournal:surchem:20150814_exp51a_635.png"><img align="left" alt="635 nm" class="medialeft" src="https://static.igem.org/mediawiki/2015/5/5b/Freiburg_labjournal-surchem-20150814_exp51a_635.png" title="635 nm" width="100"/></a>
</p>
</div>
<!-- EDIT167 SECTION "2015.08.14" [64918-65282] -->
<h3 class="sectionedit168">2015.08.15</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> DNA was let dry in</div>
</li>
<li class="level1"><div class="li"> slide 51b was washed with H<sub>2</sub>O and dried with wafergun</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> measured in Microarray Scanner for Cy5</div>
</li>
<li class="level1"><div class="li"> 3 spots could be (hardly) seen with the eye. The intensity was too low to be visible on the derived image file. Background was approximately at 0.7 to 1.7. The spot was between 1.2 and 4.0.</div>
</li>
</ul>
<p>
<a class="media" href="https://static.igem.org/mediawiki/2015/4/45/Freiburg_labjournal-surchem-20150815_exp51b_532.png" title="labjournal:surchem:20150815_exp51b_532.png"><img align="left" alt="532 nm" class="medialeft" src="https://static.igem.org/mediawiki/2015/4/45/Freiburg_labjournal-surchem-20150815_exp51b_532.png" title="532 nm" width="100"/></a>
<a class="media" href="https://static.igem.org/mediawiki/2015/2/29/Freiburg_labjournal-surchem-20150815_exp51b_635.png" title="labjournal:surchem:20150815_exp51b_635.png"><img align="left" alt="635 nm" class="medialeft" src="https://static.igem.org/mediawiki/2015/2/29/Freiburg_labjournal-surchem-20150815_exp51b_635.png" title="635 nm" width="100"/></a>
</p>
</div>
<!-- EDIT168 SECTION "2015.08.15" [65283-65798] -->
<h3 class="sectionedit169">2015.08.17</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 15 µl of cell-free expression mix (EasyExpress (Kubick) of AG Roth) were added to each flow cell</div>
</li>
<li class="level1"><div class="li"> incubation for 20 min at 37°C</div>
</li>
<li class="level1"><div class="li"> fluorescence was measured in microscope of AG Eimer</div>
</li>
<li class="level1"><div class="li"> no expressed GFP could be detected</div>
</li>
</ul>
<p>
<a class="media" href="https://static.igem.org/mediawiki/2015/5/56/Freiburg_labjournal-surchem-20150817_exp51a.png" title="labjournal:surchem:20150817_exp51a.png"><img align="left" alt="51a" class="medialeft" src="https://static.igem.org/mediawiki/2015/5/56/Freiburg_labjournal-surchem-20150817_exp51a.png" title="51a" width="100"/></a>
<a class="media" href="https://static.igem.org/mediawiki/2015/3/39/Freiburg_labjournal-surchem-20150817_exp51b.png" title="labjournal:surchem:20150817_exp51b.png"><img align="left" alt="51b" class="medialeft" src="https://static.igem.org/mediawiki/2015/3/39/Freiburg_labjournal-surchem-20150817_exp51b.png" title="51b" width="100"/></a>
</p>
</div>
<!-- EDIT169 SECTION "2015.08.17" [65799-66187] -->
<h2 class="sectionedit170">Experiment 50c: Ni-NTA slides for iRIf - rabbit serum</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT170 SECTION "Experiment 50c: Ni-NTA slides for iRIf - rabbit serum" [66188-66375] -->
<h3 class="sectionedit171">2015.08.11</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 1 Ni-NTA slides for measurement in iRIf</div>
</li>
<li class="level1"><div class="li"> slide-#: 424</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> washed 1 iRIf slides</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 80 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 1 h</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 5 h</div>
</li>
<li class="level1"><div class="li"> stored slides at 4 °C o/n</div>
</li>
</ul>
</div>
<!-- EDIT171 SECTION "2015.08.11" [66376-66661] -->
<h3 class="sectionedit172">2015.08.12</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> sandwiched slides with 60 µL NTA-solution per sandwich and incubated o/n at 4 °C</div>
</li>
</ul>
</div>
<!-- EDIT172 SECTION "2015.08.12" [66662-66798] -->
<h3 class="sectionedit173">2015.08.13</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocked slides in APTES/PDITC blocking solution for 1 h</div>
</li>
<li class="level1"><div class="li"> washed 2x with PBS</div>
</li>
<li class="level1"><div class="li"> incubated slides for 1h 15 min in NiSO&lt;Sub&gt;4&lt;/Sub&gt;-solution</div>
</li>
<li class="level1"><div class="li"> washed slides 2x with PBS</div>
</li>
<li class="level1"><div class="li"> washed slides with 1/10 diluted PBS</div>
</li>
<li class="level1"><div class="li"> stored slides for later use under N<sub>2</sub>-atmosphere at 4 °C</div>
</li>
</ul>
</div>
<!-- EDIT173 SECTION "2015.08.13" [66799-67137] -->
<h3 class="sectionedit174">2015.08.18</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 3 µL of cell free expressed GFP, GFP Lysate, GFP desalt, bBSA, BSA and mCherry were spotted.</div>
</li>
</ul>
<p>
spotting pattern:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<div class="table sectionedit175"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1-2 </td><td class="col1">His-GFP (cellfree expressed))</td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0"> 3 </td><td class="col1"> His-GFP desalt. </td><td class="col2"> 0.2 mg/ml </td>
</tr>
<tr class="row3">
<td class="col0"> 4-5 </td><td class="col1">neg. control (cellfree Expression without DNA)</td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0"> 6-7 </td><td class="col1"> His-mCherry lysate </td><td class="col2"> 1:10 </td>
</tr>
<tr class="row5">
<td class="col0"> 8-9 </td><td class="col1"> His-GFP lysate </td><td class="col2"> 1:10 </td>
</tr>
<tr class="row6">
<td class="col0"> 10 </td><td class="col1">BSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
<tr class="row7">
<td class="col0"> 11 </td><td class="col1">bBSA</td><td class="col2"> 0.2 mg/ml </td>
</tr>
</table></div>
<!-- EDIT175 TABLE [67364-67646] -->
<p>
slide (424) was measured with Serum(-) and anti-GFP
(measured together with slides from Exp. 54b)
</p>
</div>
<!-- EDIT174 SECTION "2015.08.18" [67138-67747] -->
<h3 class="sectionedit176">2015.08.19</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocked spots 2x with 10 mg/mL BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> blocked slides with 10 mg/mL BSA for 30 min</div>
</li>
<li class="level1"><div class="li"> washed slides with PBS/diluted PB + 20 mM Imidazole/diluted PBS for 10 min </div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
Flush protocol Measuring with Serum (-) [not immunized] and Serum (+) [immunized]/a-GFP:
</p>
<div class="table sectionedit177"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> # </th><th class="col2"> Flowrate </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">780</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">780</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Serum (-)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4"> * </td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">Serum (+)/a-GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4"> * / 3ug/ml </td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">anti-Rabbit</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row10">
<td class="col0">Strep Cy5</td><td class="col1">10</td><td class="col2">30</td><td class="col3">6000</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row11">
<td class="col0">Buffer</td><td class="col1">11</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT177 TABLE [68079-68415] -->
</div>
<h4>Results</h4>
<div class="level4">
</div>
<!-- EDIT176 SECTION "2015.08.19" [67748-68431] -->
<h2 class="sectionedit178">Experiment 50b: Ni-NTA slides for iRIf - Antigens and GFP lysates + purified (more HCV)</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT178 SECTION "Experiment 50b: Ni-NTA slides for iRIf - Antigens and GFP lysates + purified (more HCV)" [68432-68654] -->
<h3 class="sectionedit179">2015.08.11</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 Ni-NTA slides for measurement in iRIf</div>
</li>
<li class="level1"><div class="li"> Changed buffer: use always <strong>TBS</strong> instead of PBS (because TBS is Elution buffer) </div>
</li>
<li class="level1"><div class="li"> also use TBS for iRIf measurement</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> washed 2 iRIf slides</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 80 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 1 h</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 5 h</div>
</li>
<li class="level1"><div class="li"> stored slides at 4 °C o/n</div>
</li>
</ul>
</div>
<!-- EDIT179 SECTION "2015.08.11" [68655-69048] -->
<h3 class="sectionedit180">2015.08.12</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> sandwiched slides with 60 µL NTA-solution per sandwich and incubated o/n at 4 °C</div>
</li>
</ul>
</div>
<!-- EDIT180 SECTION "2015.08.12" [69049-69185] -->
<h3 class="sectionedit181">2015.08.13</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocked slides in APTES/PDITC blocking solution for 1 h</div>
</li>
<li class="level1"><div class="li"> washed 2x with PBS</div>
</li>
<li class="level1"><div class="li"> incubated slides for 1h 15 min in NiSO<sub>4</sub> -solution</div>
</li>
<li class="level1"><div class="li"> washed slides 2x with PBS</div>
</li>
<li class="level1"><div class="li"> washed slides with 1/10 diluted PBS</div>
</li>
<li class="level1"><div class="li"> spotted antigens were diluted in TBS</div>
</li>
<li class="level1"><div class="li"> spotted slides <strong>500</strong> and <strong>501</strong></div>
</li>
</ul>
<p>
Spotting:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<div class="table sectionedit182"><table class="inline">
<tr class="row0">
<th class="col0"> Spot </th><th class="col1"> Protein </th><th class="col2"> Number of Spots </th><th class="col3"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-2</td><td class="col1"> pET_1003 (HCV) lysate </td><td class="col2"> 2 </td><td class="col3">~0.08 mg/ml</td>
</tr>
<tr class="row2">
<td class="col0">3-4</td><td class="col1"> pET_1003 (HCV) desalt </td><td class="col2"> 2 </td><td class="col3">~0.33 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">5</td><td class="col1"> pET_1703 (HIV) lysate </td><td class="col2"> 1 </td><td class="col3">~0.15 mg/ml</td>
</tr>
<tr class="row4">
<td class="col0">6</td><td class="col1"> pET_1703 (HIV) desalt </td><td class="col2"> 1 </td><td class="col3">~0.62 mg/ml</td>
</tr>
<tr class="row5">
<td class="col0">7</td><td class="col1"> His-GFP (Max)</td><td class="col2"> 1 </td><td class="col3">0.5 mg/ml</td>
</tr>
<tr class="row6">
<td class="col0">8</td><td class="col1"> His-GFP lysate </td><td class="col2"> 1 </td><td class="col3">~0.2 mg/ml</td>
</tr>
<tr class="row7">
<td class="col0">9</td><td class="col1"> bBSA </td><td class="col2"> 1 </td><td class="col3">0.2 mg/ml</td>
</tr>
<tr class="row8">
<td class="col0">10-11</td><td class="col1"> His-GFP aufgereinigt </td><td class="col2"> 2 </td><td class="col3">~0.5 mg/ml</td>
</tr>
</table></div>
<!-- EDIT182 TABLE [69609-69981] -->
</div>
<!-- EDIT181 SECTION "2015.08.13" [69186-69982] -->
<h3 class="sectionedit183">2015.08.14</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA in <strong>TBS</strong> for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked with 10 mg/mL BSA in <strong>TBS</strong> for 30 min</div>
</li>
<li class="level1"><div class="li"> slides were washed with <strong>TBS</strong></div>
</li>
<li class="level1"><div class="li"> slides were washed with 1/10 diluted <strong>TBS</strong> + 20 mM imidazole</div>
</li>
<li class="level1"><div class="li"> slides were washed with 1/10 diluted <strong>TBS</strong></div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
slide 500 was measured in new setup<br/>

slide 501 could not be measured, air bubbles appeared every time the slide was flushed –&gt; surface must be really hydrophobic. Maybe slide was destroyed during tests with PDMS-stamps.
</p>
<div class="table sectionedit184"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer (TBS!!!)</td><td class="col1">1</td><td class="col2">60</td><td class="col3">720</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0 leftalign">BSA	</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Anti-HIV (polyclonal,rabbit)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">5</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">Anti-HCV (polyclonal,rabbit)</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">7</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">Anti-Rabbit (from mouse)</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row10">
<td class="col0">Anti-HIV (monoclonal,mouse)</td><td class="col1">10</td><td class="col2">30</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row11">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">11</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row12">
<td class="col0">Anti-HCV (monoclonal,mouse)</td><td class="col1">12</td><td class="col2">30</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row13">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">13</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row14">
<td class="col0">Anti-His</td><td class="col1">14</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row15">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">15</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row16">
<td class="col0">Anti-GFP (biotinylated, goat)</td><td class="col1">16</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row17">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">17</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row18">
<td class="col0">StrepCy5</td><td class="col1">18</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row19">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">19</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT184 TABLE [70551-71299] -->
</div>
<h4>Results</h4>
<div class="level4">
</div>
<!-- EDIT183 SECTION "2015.08.14" [69983-71315] -->
<h2 class="sectionedit185">Experiment 50a: Ni-NTA slides for iRIf - Antigens and GFP lysates + purified (more HIV)</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT185 SECTION "Experiment 50a: Ni-NTA slides for iRIf - Antigens and GFP lysates + purified (more HIV)" [71316-71538] -->
<h3 class="sectionedit186">2015.08.11</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 Ni-NTA slides for measurement in iRIf</div>
</li>
<li class="level1"><div class="li"> Changed buffer: use always <strong>TBS</strong> instead of PBS (because TBS is Elution buffer) </div>
</li>
<li class="level1"><div class="li"> also use TBS for iRIf measurement</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> washed 2 iRIf slides</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 80 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 1 h</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 5 h</div>
</li>
<li class="level1"><div class="li"> stored slides at 4 °C o/n</div>
</li>
</ul>
</div>
<!-- EDIT186 SECTION "2015.08.11" [71539-71932] -->
<h3 class="sectionedit187">2015.08.12</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> sandwiched slides with 60 µL NTA-solution per sandwich and incubated o/n at 4 °C</div>
</li>
</ul>
</div>
<!-- EDIT187 SECTION "2015.08.12" [71933-72069] -->
<h3 class="sectionedit188">2015.08.13</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocked slides in APTES/PDITC blocking solution for 1 h</div>
</li>
<li class="level1"><div class="li"> washed 2x with PBS</div>
</li>
<li class="level1"><div class="li"> incubated slides for 1h 15 min in NiSO<sub>4</sub>-solution</div>
</li>
<li class="level1"><div class="li"> washed slides 2x with PBS</div>
</li>
<li class="level1"><div class="li"> washed slides with 1/10 diluted PBS</div>
</li>
<li class="level1"><div class="li"> antigenes were diluted in TBS</div>
</li>
<li class="level1"><div class="li"> spotted slide <strong>216</strong> and <strong>467</strong></div>
</li>
</ul>
<p>
Spotting:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<div class="table sectionedit189"><table class="inline">
<tr class="row0">
<th class="col0"> Spot </th><th class="col1"> Protein </th><th class="col2"> Number of Spots </th><th class="col3"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-2</td><td class="col1"> pET_1703 (HIV) lysate </td><td class="col2"> 2 </td><td class="col3">~0.15 mg/ml</td>
</tr>
<tr class="row2">
<td class="col0">3-4</td><td class="col1"> pET_1703 (HIV) desalt</td><td class="col2"> 2 </td><td class="col3">~0.62 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">5</td><td class="col1"> pET_1003 (HCV) lysate </td><td class="col2"> 1 </td><td class="col3">~0.08 mg/ml</td>
</tr>
<tr class="row4">
<td class="col0">6</td><td class="col1"> pET_1003 (HCV) desalt</td><td class="col2"> 1 </td><td class="col3">~0.33 mg/ml</td>
</tr>
<tr class="row5">
<td class="col0">7</td><td class="col1"> His-GFP (Max)</td><td class="col2"> 1 </td><td class="col3">0.5 mg/ml</td>
</tr>
<tr class="row6">
<td class="col0">8</td><td class="col1"> His-GFP desalt </td><td class="col2"> 1 </td><td class="col3">~0.5 mg/ml</td>
</tr>
<tr class="row7">
<td class="col0">9</td><td class="col1"> bBSA </td><td class="col2"> 1 </td><td class="col3">0.2 mg/ml</td>
</tr>
<tr class="row8">
<td class="col0">10-11</td><td class="col1"> His-GFP lysate </td><td class="col2"> 2 </td><td class="col3">~0.2 mg/ml</td>
</tr>
</table></div>
<!-- EDIT189 TABLE [72483-72847] -->
</div>
<!-- EDIT188 SECTION "2015.08.13" [72070-72848] -->
<h3 class="sectionedit190">2015.08.14</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA in PBS for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked with 10 mg/mL BSA in <strong>TBS</strong> for 30 min</div>
</li>
<li class="level1"><div class="li"> slides were washed with <strong>TBS</strong></div>
</li>
<li class="level1"><div class="li"> slides were washed with 1/10 diluted <strong>TBS</strong> + 20 mM imidazole</div>
</li>
<li class="level1"><div class="li"> slides were washed with 1/10 diluted <strong>TBS</strong></div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
216 was measured in old Setup´<br/>

467 was measured in new Setup
</p>
<div class="table sectionedit191"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer (TBS!!!)</td><td class="col1">1</td><td class="col2">60</td><td class="col3">720</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0 leftalign">BSA	</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Anti-HIV (polyclonal,rabbit)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">5</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">Anti-HCV (polyclonal,rabbit)</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">7</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">Anti-Rabbit (from mouse)</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row10">
<td class="col0">Anti-HIV (monoclonal,mouse)</td><td class="col1">10</td><td class="col2">30</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row11">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">11</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row12">
<td class="col0">Anti-HCV (monoclonal,mouse)</td><td class="col1">12</td><td class="col2">30</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row13">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">13</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row14">
<td class="col0">Anti-Mouse</td><td class="col1">14</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row15">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">15</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row16">
<td class="col0">Anti-GFP (biotinylated, goat)</td><td class="col1">16</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row17">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">17</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row18">
<td class="col0">StrepCy5</td><td class="col1">18</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row19">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">19</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT191 TABLE [73256-74006] -->
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/3/31/Freiburg_labjournal-roi_20150815_24.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:roi_20150815_24.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 24: Selection of ROIs: 1 HIV spot (spot 2) and one HCV spot (spot 3) were excluded due to high amounts of salt on it</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/71/Freiburg_labjournal-binding_curve_20150815_24.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:binding_curve_20150815_24.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 24: Binding curves</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/f1/Freiburg_labjournal-roi_20150815_502.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:roi_20150815_502.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 502: Selection of ROIs</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/c/c0/Freiburg_labjournal-binding_curve_20150815_502_complete.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:binding_curve_20150815_502_complete.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 502: Binding curves for all spots</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/d/d6/Freiburg_labjournal-binding_curve_20150815_502_salm1.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:binding_curve_20150815_502_salm1.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 502: Binding curve (second Salmonella spot excluded)</div></div></div><hr/>
<p>
Results: We have specific antibody/antigen binding for Salmonella! (a-HIV/HCV do not bind…) 
Slide 502: During measurement within the first buffer step/blocking step most of the second Salmonella spot was washed away.
Related to this the spot behaves strange.
Slide 24: Also anti-salmonella lysate was tested. Here we see (as expected) stronger unspecific binding at other spots (nevertheless greatest binding for the salmonella spots).
The unspecific/transient binding proteins get mostly washed away during the following buffer step.
</p>
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/c/c4/Freiburg_labjournal-qp_24_a-salm_a-gfp.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:qp_24_a-salm_a-gfp.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 24: Quotient picture (before a-Salmonella / after a-Salmonella + a-GFP)</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/f3/Freiburg_labjournal-qp_502_a-salm_a-gfp.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:qp_502_a-salm_a-gfp.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 502: Quotient picture (before a-Salmonella / after a-Salmonella + a-GFP)</div></div></div>
</div>
<!-- EDIT190 SECTION "2015.08.14" [72849-75399] -->
<h2 class="sectionedit192">Experiment 49c: PDITC slides for iRIf</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT192 SECTION "Experiment 49c: PDITC slides for iRIf" [75400-75572] -->
<h3 class="sectionedit193">2015.08.11</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 1 PDITC slides for measurement in iRIf</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> washed 1 iRIf slides</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 80 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 1 h</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 5 h</div>
</li>
<li class="level1"><div class="li"> stored slides at 4 °C under N<sub>2</sub>-atmosphere</div>
</li>
</ul>
</div>
<!-- EDIT193 SECTION "2015.08.11" [75573-75867] -->
<h3 class="sectionedit194">2015.08.13</h3>
<div class="level3">
<ul>
<li class="level1"><div class="li"> used slides for measurement in own device –&gt; Exp. 52</div>
</li>
</ul>
</div>
<!-- EDIT194 SECTION "2015.08.13" [75868-75945] -->
<h2 class="sectionedit195">Experiment 49b: PDITC slides for iRIf - Antigens</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT195 SECTION "Experiment 49b: PDITC slides for iRIf - Antigens" [75946-76129] -->
<h3 class="sectionedit196">2015.08.11</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 PDITC slides for measurement in iRIf</div>
</li>
<li class="level1"><div class="li"> Changed buffer: use always <strong>TBS</strong> instead of PBS (because TBS is Elution buffer) </div>
</li>
<li class="level1"><div class="li"> also use TBS for iRIf measurement</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> washed 2 iRIf slides</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 80 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 1 h</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 5 h</div>
</li>
<li class="level1"><div class="li"> stored slides at 4 °C under N<sub>2</sub>-atmosphere</div>
</li>
</ul>
</div>
<!-- EDIT196 SECTION "2015.08.11" [76130-76548] -->
<h3 class="sectionedit197">2015.08.13</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spotted slides <strong>502</strong> and <strong>024</strong></div>
</li>
<li class="level1"><div class="li"> incubated at 4°C o/n</div>
</li>
</ul>
<p>
Spotting:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<div class="table sectionedit198"><table class="inline">
<tr class="row0">
<th class="col0"> Spot </th><th class="col1"> Protein </th><th class="col2"> Number of spots </th><th class="col3"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-2</td><td class="col1"> pET_1003 (HCV) desalt </td><td class="col2"> 2 </td><td class="col3"> ~0.33 mg/ml </td>
</tr>
<tr class="row2">
<td class="col0">3-4</td><td class="col1"> pET_1703 (HIV) desalt</td><td class="col2"> 2 </td><td class="col3"> ~0.62 mg/ml </td>
</tr>
<tr class="row3">
<td class="col0">5-6</td><td class="col1"> pIG_1501 (Salmonella) desalt</td><td class="col2"> 2 </td><td class="col3"> ~0.62mg/ml </td>
</tr>
<tr class="row4">
<td class="col0">7</td><td class="col1"> GFP-His (Max) </td><td class="col2"> 1 </td><td class="col3"> 0.5 mg/ml</td>
</tr>
<tr class="row5">
<td class="col0">8</td><td class="col1"> GFP-His desalt </td><td class="col2"> 1 </td><td class="col3"> 0.5 mg/ml</td>
</tr>
<tr class="row6">
<td class="col0">9</td><td class="col1"> GFP-His lysate </td><td class="col2"> 1 </td><td class="col3"> ~0.2 mg/ml</td>
</tr>
<tr class="row7">
<td class="col0">10</td><td class="col1"> bBSA </td><td class="col2"> 1 </td><td class="col3"> 0.2 mg/ml</td>
</tr>
<tr class="row8">
<td class="col0">11</td><td class="col1"> BSA </td><td class="col2"> 1 </td><td class="col3"> 0.2 mg/ml </td>
</tr>
</table></div>
<!-- EDIT198 TABLE [76736-77098] -->
</div>
<!-- EDIT197 SECTION "2015.08.13" [76549-77099] -->
<h3 class="sectionedit199">2015.08.13</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA in PBS (should have used TBS) for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked with 10 mg/mL BSA in PBS for 30 min</div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<div class="table sectionedit200"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer (TBS!!!)</td><td class="col1">1</td><td class="col2">60</td><td class="col3">720</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0 leftalign">BSA	</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Anti-HIV (polyclonal,rabbit)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">5</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">Anti-HCV (polyclonal,rabbit)</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">Anti-Rabbit (from mouse)</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row10">
<td class="col0">Anti-HIV (monoclonal,mouse)</td><td class="col1">10</td><td class="col2">30</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row11">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">11</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row12">
<td class="col0">Anti-HCV (monoclonal,mouse)</td><td class="col1">12</td><td class="col2">30</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row13">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">13</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row14">
<td class="col0">Anti-Mouse</td><td class="col1">14</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row15">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">15</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row16">
<td class="col0">Anti-GFP (biotinylated, goat)</td><td class="col1">16</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row17">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">17</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row18">
<td class="col0">Anti-Salmonella (13)</td><td class="col1">18</td><td class="col2">30</td><td class="col3">600</td><td class="col4">~50-80 ug/ml</td>
</tr>
<tr class="row19">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">19</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row20">
<td class="col0">StrepCy5</td><td class="col1">20</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row21">
<td class="col0">Buffer	(TBS!!!)</td><td class="col1">21</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT200 TABLE [77307-78134] -->
</div>
<!-- EDIT199 SECTION "2015.08.13" [77100-78135] -->
<h2 class="sectionedit201">Experiment 49a: PDITC slides for iRIf - bBSA/BSA for new device</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT201 SECTION "Experiment 49a: PDITC slides for iRIf - bBSA/BSA for new device" [78136-78334] -->
<h3 class="sectionedit202">2015.08.11</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 PDITC slides for measurement in iRIf</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> washed iRIf slides</div>
</li>
<li class="level1"><div class="li"> Plasma activation: 80 L/h, 5 min</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 1 h</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 5 h</div>
</li>
<li class="level1"><div class="li"> spotted 2 slides (426, 253) with bBSA/BSA for new device</div>
</li>
</ul>
</div>
<!-- EDIT202 SECTION "2015.08.11" [78335-78630] -->
<h3 class="sectionedit203">2015.08.12</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocked spots 2x with 10 mg/mL BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> blocked slides with 10 mg/mL BSA for 30 min</div>
</li>
<li class="level1"><div class="li"> washed slides with ddH<sub>2</sub>O and dried with wafer gun</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
</div>
<!-- EDIT203 SECTION "2015.08.12" [78631-78860] -->
<h2 class="sectionedit204">Experiment 48: DNA on PDMS</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used : <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/PDMS_PDITC" title="protocols:pdms_pditc">PDMS-PDITC</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a></div>
</li>
</ul>
</div>
<!-- EDIT204 SECTION "Experiment 48: DNA on PDMS" [78861-79008] -->
<h3 class="sectionedit205">2015.08.11</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> immobilize Cy5-labeled DNA on PDMS slides</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> used 2 flow chamber slides with 3 holes</div>
</li>
<li class="level1"><div class="li"> washed PDMS slides with Ethanol and H<sub>2</sub>O</div>
</li>
<li class="level1"><div class="li"> used just 200 µL APTES in <strong>Acetone</strong> for APTES solution instead of 1 mL APTES in Ethanol </div>
</li>
<li class="level1"><div class="li"> activated slides in  Plasma Generator with 40 L/h for 1 min</div>
</li>
<li class="level1"><div class="li"> incubated slides in APTES solution for 2 days+</div>
</li>
</ul>
</div>
<!-- EDIT205 SECTION "2015.08.11" [79009-79436] -->
<h3 class="sectionedit206">2015.08.13</h3>
<div class="level3">
<ul>
<li class="level1"><div class="li"> each slide was washed with ethanol</div>
</li>
<li class="level1"><div class="li"> a 5 min washing step in Ethanol followed</div>
</li>
<li class="level1"><div class="li"> after drying with wafergun slides were baked in oven for 90 min at 70°C</div>
</li>
<li class="level1"><div class="li"> slides were cooled down with wafergun</div>
</li>
<li class="level1"><div class="li"> incubation in PDITC for 5h40min at RT</div>
</li>
<li class="level1"><div class="li"> filled slideholder with EtOH and discarded it immediately</div>
</li>
<li class="level1"><div class="li"> washed 2x 5 min with EtOH in slideholder</div>
</li>
<li class="level1"><div class="li"> filled slideholder again with EtOH and discarded it immediately</div>
</li>
<li class="level1"><div class="li"> dried with wafergun</div>
</li>
<li class="level1"><div class="li"> 1 slide (a) was spotted with 1 µl spots and incubated over night</div>
</li>
<li class="level1"><div class="li"> sample 1 was diluted 1:4,64 (original oncentration 116 ng/µl) in NaPi buffer 150 mM</div>
</li>
</ul>
<p>
spotting pattern 48a:
<a class="media" href="https://static.igem.org/mediawiki/2015/5/5b/Freiburg_labjournal-spotting_pattern_pdms_1.png" title="labjournal:spotting_pattern_pdms_1.png"><img align="left" alt="" class="medialeft" src="https://static.igem.org/mediawiki/2015/5/5b/Freiburg_labjournal-spotting_pattern_pdms_1.png" width="200"/></a>
</p>
<div class="table sectionedit207"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1</td><td class="col1">His-tYFP-Spy (lina104) (top 0.5 µl, bottom spot 1 µl)</td><td class="col2">25 ng/µl</td>
</tr>
<tr class="row2">
<td class="col0">2</td><td class="col1">His-tYFP-Halo (lina105)</td><td class="col2">20 ng/µl</td>
</tr>
<tr class="row3">
<td class="col0">3</td><td class="col1">HA-GFP-His<sub>6</sub>-His<sub>6</sub></td><td class="col2">28.7 ng/µl</td>
</tr>
<tr class="row4">
<td class="col0">4</td><td class="col1">Control by Tobi: HA-GFP-Halo-His</td><td class="col2">25 ng/µl</td>
</tr>
</table></div>
<!-- EDIT207 TABLE [80135-80373] --><ul>
<li class="level1"><div class="li"> slide (b) was spotted with 3 µl spots and incubated for 2 nights at room temperature</div>
</li>
<li class="level1"><div class="li"> sample 1 was diluted 1:4,64 (original concentration 116 ng/µl) in NaPi buffer 150 mM</div>
</li>
</ul>
<p>
spotting pattern 48b:
<a class="media" href="https://static.igem.org/mediawiki/2015/5/5b/Freiburg_labjournal-spotting_pattern_pdms_1.png" title="labjournal:spotting_pattern_pdms_1.png"><img align="left" alt="" class="medialeft" src="https://static.igem.org/mediawiki/2015/5/5b/Freiburg_labjournal-spotting_pattern_pdms_1.png" width="200"/></a>
</p>
<div class="table sectionedit208"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1</td><td class="col1">His-tYFP-Spy (lina104)</td><td class="col2">25 ng/µl</td>
</tr>
<tr class="row2">
<td class="col0">2</td><td class="col1">His-tYFP-Halo (lina105)</td><td class="col2">20 ng/µl</td>
</tr>
<tr class="row3">
<td class="col0">3</td><td class="col1">HA-GFP-His<sub>6</sub>-His<sub>6</sub></td><td class="col2">28.7 ng/µl</td>
</tr>
<tr class="row4">
<td class="col0">4</td><td class="col1">Control by Tobi: HA-GFP-Halo-HisHis</td><td class="col2">25 ng/µl</td>
</tr>
</table></div>
<!-- EDIT208 TABLE [80629-80837] -->
</div>
<!-- EDIT206 SECTION "2015.08.13" [79437-80838] -->
<h3 class="sectionedit209">2015.08.14</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> DNA was let dry in</div>
</li>
<li class="level1"><div class="li"> slide 48a was washed with H<sub>2</sub>O and dried with wafergun</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slide48a was measured in Microarray Scanner for Cy5</div>
</li>
<li class="level1"><div class="li"> no spots could be seen  </div>
</li>
</ul>
<p>
<a class="media" href="https://static.igem.org/mediawiki/2015/7/7b/Freiburg_labjournal-surchem-20150814_exp48a_532.png" title="labjournal:surchem:20150814_exp48a_532.png"><img align="left" alt="532 nm" class="medialeft" src="https://static.igem.org/mediawiki/2015/7/7b/Freiburg_labjournal-surchem-20150814_exp48a_532.png" title="532 nm" width="100"/></a>
<a class="media" href="https://static.igem.org/mediawiki/2015/2/23/Freiburg_labjournal-surchem-20150814_exp48a_635.png" title="labjournal:surchem:20150814_exp48a_635.png"><img align="left" alt="635 nm" class="medialeft" src="https://static.igem.org/mediawiki/2015/2/23/Freiburg_labjournal-surchem-20150814_exp48a_635.png" title="635 nm" width="100"/></a>
</p>
</div>
<!-- EDIT209 SECTION "2015.08.14" [80839-81202] -->
<h3 class="sectionedit210">2015.08.15</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> DNA was let dry in</div>
</li>
<li class="level1"><div class="li"> slide 48b was washed with H<sub>2</sub>O and dried with wafergun</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slide48b was measured in Microarray Scanner for Cy5</div>
</li>
<li class="level1"><div class="li"> no spots could be seen  </div>
</li>
</ul>
<p>
<a class="media" href="https://static.igem.org/mediawiki/2015/1/1c/Freiburg_labjournal-surchem-20150815_exp48b_532.png" title="labjournal:surchem:20150815_exp48b_532.png"><img align="left" alt="532 nm" class="medialeft" src="https://static.igem.org/mediawiki/2015/1/1c/Freiburg_labjournal-surchem-20150815_exp48b_532.png" title="532 nm" width="100"/></a>
<a class="media" href="https://static.igem.org/mediawiki/2015/2/26/Freiburg_labjournal-surchem-20150815_exp48b_635.png" title="labjournal:surchem:20150815_exp48b_635.png"><img align="left" alt="635 nm" class="medialeft" src="https://static.igem.org/mediawiki/2015/2/26/Freiburg_labjournal-surchem-20150815_exp48b_635.png" title="635 nm" width="100"/></a>
</p>
</div>
<!-- EDIT210 SECTION "2015.08.15" [81203-81568] -->
<h2 class="sectionedit211">Experiment 47b: PDITC slides for iRIf</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT211 SECTION "Experiment 47b: PDITC slides for iRIf" [81569-81741] -->
<h3 class="sectionedit212">2015.08.09</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 1 PDITC iRIf slides for measurement of antigens, PhyA or bBSA in own device</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 1 PDITC slides</div>
</li>
<li class="level1"><div class="li"> plasma activation: 80 L/h</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
<li class="level1"><div class="li"> slides were stored in slide holder under N<sub>2</sub>-atmosphere</div>
</li>
</ul>
</div>
<!-- EDIT212 SECTION "2015.08.09" [81742-82084] -->
<h3 class="sectionedit213">2015.08.13</h3>
<div class="level3">
<ul>
<li class="level1"><div class="li"> used slides for measurement in own device –&gt; Exp. 52</div>
</li>
</ul>
</div>
<!-- EDIT213 SECTION "2015.08.13" [82085-82164] -->
<h2 class="sectionedit214">Experiment 47a: PDITC slides for iRIf - PhyA Testing</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT214 SECTION "Experiment 47a: PDITC slides for iRIf - PhyA Testing" [82165-82352] -->
<h3 class="sectionedit215">2015.08.09</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 PDITC iRIf slides for measurement of antigens, PhyA or bBSA in own device</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 2 PDITC slides</div>
</li>
<li class="level1"><div class="li"> plasma activation: 80 L/h</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
<li class="level1"><div class="li"> slides were stored in slide holder under N<sub>2</sub>-atmosphere</div>
</li>
</ul>
</div>
<!-- EDIT215 SECTION "2015.08.09" [82353-82695] -->
<h3 class="sectionedit216">2015.08.10</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spotted 3 µL of different PhyAs and controls on 2 slides</div>
</li>
</ul>
<p>
Spotting:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<div class="table sectionedit217"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> Elution no. </th><th class="col3"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1-2 </td><td class="col1">PhyA-GFP </td><td class="col2 leftalign"> </td><td class="col3 leftalign"> </td>
</tr>
<tr class="row2">
<td class="col0"> 3-4 </td><td class="col1">His-PhyA-GST selbst aufgereinigt</td><td class="col2"> 2 </td><td class="col3"> 6.6 mg/ml → 1:10 </td>
</tr>
<tr class="row3">
<td class="col0"> 5 </td><td class="col1"> His-PhyA-GST in AntigenSolution (17) </td><td class="col2"> 2 </td><td class="col3 leftalign"> 1:1 (1:10 His-PhyA-GST in 5mg/ml BSA : AntigenSolution (1:5)  </td>
</tr>
<tr class="row4">
<td class="col0"> 6 </td><td class="col1">GFP (Max) </td><td class="col2 leftalign"> </td><td class="col3"> 1 mg/ml </td>
</tr>
<tr class="row5">
<td class="col0"> 7 </td><td class="col1">GFP-Lysate-NT </td><td class="col2 leftalign"> </td><td class="col3"></td>
</tr>
<tr class="row6">
<td class="col0"> 8 </td><td class="col1">His-GFP-Halo (Konstrukt von AG-Roth) - selbst aufgereinigt </td><td class="col2"> ? </td><td class="col3 leftalign"> </td>
</tr>
<tr class="row7">
<td class="col0"> 9 </td><td class="col1">bBSA </td><td class="col2 leftalign"> </td><td class="col3"> 0.2 mg/ml </td>
</tr>
<tr class="row8">
<td class="col0"> 10 </td><td class="col1"> GFP (Max) in AntigenSolution (17)</td><td class="col2"> </td><td class="col3"> 1:1 (0.2 mg/ml GFP in 5mg/ml BSA : AntigenSolution (1:5))</td>
</tr>
<tr class="row9">
<td class="col0"> 11 </td><td class="col1"> GFP (Max) in Elutionspuffer </td><td class="col2"> </td><td class="col3"> 1:1 (0.2 mg/ml GFP in 5mg/ml BSA : Elutionsbuffer) </td>
</tr>
</table></div>
<!-- EDIT217 TABLE [82883-83490] -->
</div>
<!-- EDIT216 SECTION "2015.08.10" [82696-83491] -->
<h3 class="sectionedit218">2015.08.11</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> sildes were blocked with 10 mg/mL BSA for 30 min</div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
slide 012 was measured, the other slide is broken and could not be measured.
</p>
<div class="table sectionedit219"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">780</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">420</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">anti-phyA (N-term)</td><td class="col1">4</td><td class="col2">40</td><td class="col3">450</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">anti-phyA (rabbit, polyclonal)</td><td class="col1">6</td><td class="col2">40</td><td class="col3">450</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">anti-GFP (goat, biotinylated)</td><td class="col1">8</td><td class="col2">40</td><td class="col3">450</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row10">
<td class="col0">anti-GST</td><td class="col1">10</td><td class="col2">40</td><td class="col3">450</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row11">
<td class="col0">Buffer</td><td class="col1">11</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row12">
<td class="col0">anti-Rabbit (mouse)</td><td class="col1">12</td><td class="col2">40</td><td class="col3">450</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row13">
<td class="col0">Buffer</td><td class="col1">13</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row14">
<td class="col0">anti-goat</td><td class="col1">14</td><td class="col2">40</td><td class="col3">450</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row15">
<td class="col0">Buffer</td><td class="col1">15</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
<tr class="row16">
<td class="col0">StrepCy5</td><td class="col1">16</td><td class="col2">40</td><td class="col3">450</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row17">
<td class="col0">Buffer</td><td class="col1">17</td><td class="col2">60</td><td class="col3">300</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT219 TABLE [83747-84299] -->
</div>
<h4>Results</h4>
<div class="level4">
</div>
<!-- EDIT218 SECTION "2015.08.11" [83492-84318] -->
<h2 class="sectionedit220">Experiment 46c: Ni-NTA slides for iRIf</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT220 SECTION "Experiment 46c: Ni-NTA slides for iRIf" [84319-84492] -->
<h3 class="sectionedit221">2015.08.09</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 Ni-NTA slides for measurement of cell free expressed Proteins (GFP/tYFP) or antigen lysate</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 2 PDITC slides</div>
</li>
<li class="level1"><div class="li"> plasma activation: 80 L/h</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 45 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2h</div>
</li>
<li class="level1"><div class="li"> slides were sandwiched with NTA solution, 1 slide was sandwiched with a normal glass slide</div>
</li>
</ul>
</div>
<!-- EDIT221 SECTION "2015.08.09" [84493-84876] -->
<h3 class="sectionedit222">2015.08.10</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slides were incubated in blocking solution  for 1 h</div>
</li>
<li class="level1"><div class="li"> slides were incubated in Nickel sulfate solution for 1 h</div>
</li>
<li class="level1"><div class="li"> slides were washed 2x in PBS and 1x in diluted PBS</div>
</li>
<li class="level1"><div class="li"> slides were dried and stored under N<sub>2</sub>-atmosphere</div>
</li>
</ul>
</div>
<!-- EDIT222 SECTION "2015.08.10" [84877-85162] -->
<h3 class="sectionedit223">2015.08.16</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spotted slides <strong>303</strong> and <strong>429</strong> with samples from cell-free</div>
</li>
<li class="level1"><div class="li"> incubated at 4°C o/n </div>
</li>
</ul>
<p>
Spotting (Slide 303):
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<div class="table sectionedit224"><table class="inline">
<tr class="row0">
<th class="col0"> Spot </th><th class="col1"> Protein </th><th class="col2"> Number of spots </th><th class="col3"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-2</td><td class="col1"> BK foil 1 </td><td class="col2"> 2 </td><td class="col3"> unknown </td>
</tr>
<tr class="row2">
<td class="col0">3-4</td><td class="col1"> neg foil 1 </td><td class="col2"> 2 </td><td class="col3"> unknown </td>
</tr>
<tr class="row3">
<td class="col0">5-6</td><td class="col1"> BK 1 </td><td class="col2"> 2 </td><td class="col3"> unknown </td>
</tr>
<tr class="row4">
<td class="col0">7</td><td class="col1"> GFP-His (Max) </td><td class="col2"> 1 </td><td class="col3"> 0.5 mg/ml</td>
</tr>
<tr class="row5">
<td class="col0">8</td><td class="col1"> GFP-His desalt </td><td class="col2"> 1 </td><td class="col3"> 0.5 mg/ml</td>
</tr>
<tr class="row6">
<td class="col0">9</td><td class="col1"> bBSA </td><td class="col2"> 1 </td><td class="col3"> 0.2 mg/ml</td>
</tr>
<tr class="row7">
<td class="col0">10-11</td><td class="col1 leftalign"> neg 1  </td><td class="col2"> 1 </td><td class="col3"> unknown </td>
</tr>
</table></div>
<!-- EDIT224 TABLE [85391-85666] -->
<p>
Spotting (Slide 429):
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<div class="table sectionedit225"><table class="inline">
<tr class="row0">
<th class="col0"> Spot </th><th class="col1"> Protein </th><th class="col2"> Number of spots </th><th class="col3"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-2</td><td class="col1"> BK foil 2 </td><td class="col2"> 2 </td><td class="col3"> unknown </td>
</tr>
<tr class="row2">
<td class="col0">3-4</td><td class="col1"> neg foil </td><td class="col2">2  2 </td><td class="col3"> unknown </td>
</tr>
<tr class="row3">
<td class="col0">5-6</td><td class="col1"> BK 2 </td><td class="col2"> 2 </td><td class="col3"> unknown </td>
</tr>
<tr class="row4">
<td class="col0">7</td><td class="col1"> GFP-His (Max) </td><td class="col2"> 1 </td><td class="col3"> 0.5 mg/ml</td>
</tr>
<tr class="row5">
<td class="col0">8</td><td class="col1"> GFP-His lysate </td><td class="col2"> 1 </td><td class="col3"> ~0.2 mg/ml</td>
</tr>
<tr class="row6">
<td class="col0">9</td><td class="col1"> bBSA </td><td class="col2"> 1 </td><td class="col3"> 0.2 mg/ml</td>
</tr>
<tr class="row7">
<td class="col0">10-11</td><td class="col1"> neg 2 </td><td class="col2"> 1 </td><td class="col3"> unknown </td>
</tr>
</table></div>
<!-- EDIT225 TABLE [85754-86027] -->
</div>
<!-- EDIT223 SECTION "2015.08.16" [85163-86027] -->
<h3 class="sectionedit226">Results</h3>
<div class="level3">
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/41/Freiburg_labjournal-surchem-exp_46c_slide429.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_46c_slide429.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 429: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/6a/Freiburg_labjournal-surchem-exp_46c_slide429_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_46c_slide429_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 429: Picture obtained from fluorescence scanner. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/8/87/Freiburg_labjournal-surchem-exp_46c_slide429_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_46c_slide429_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 429: Binding curves</div></div></div><hr/>
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/f6/Freiburg_labjournal-surchem-exp_46c_slide303.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_46c_slide303.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 303: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/a/a7/Freiburg_labjournal-surchem-exp_46c_slide303_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_46c_slide303_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 303: Picture obtained from fluorescence scanner. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/d/db/Freiburg_labjournal-surchem-exp_46c_slide303-binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_46c_slide303-binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 303: Binding curves</div></div></div>
</div>
<!-- EDIT226 SECTION "Results" [86028-86752] -->
<h2 class="sectionedit227">Experiment 46b: Ni-NTA slides for iRIf - cell free expressed GFP</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT227 SECTION "Experiment 46b: Ni-NTA slides for iRIf - cell free expressed GFP" [86753-86952] -->
<h3 class="sectionedit228">2015.08.09</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 1 Ni-NTA slides for measurement of cell free expressed Proteins (GFP/tYFP) or antigen lysate</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 1 PDITC slides</div>
</li>
<li class="level1"><div class="li"> plasma activation: 80 L/h</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 45 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2h</div>
</li>
<li class="level1"><div class="li"> slides were sandwiched with NTA solution, 1 slide was sandwiched with a normal glass slide</div>
</li>
</ul>
</div>
<!-- EDIT228 SECTION "2015.08.09" [86953-87336] -->
<h3 class="sectionedit229">2015.08.10</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slides were incubated in blocking solution  for 1 h</div>
</li>
<li class="level1"><div class="li"> slides were incubated in Nickel sulfate solution for 1 h</div>
</li>
<li class="level1"><div class="li"> slides were washed 2x in PBS and 1x in diluted PBS</div>
</li>
<li class="level1"><div class="li"> slides were dried and stored under N<sub>2</sub>-atmosphere</div>
</li>
</ul>
</div>
<!-- EDIT229 SECTION "2015.08.10" [87337-87622] -->
<h3 class="sectionedit230">2015.08.13</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spotted cell-free expressed GFP and His-Lysate on slide 208</div>
</li>
<li class="level1"><div class="li"> incubation o/n at 4 °C </div>
</li>
</ul>
<p>
Spotting:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<div class="table sectionedit231"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1-3 </td><td class="col1">HA-GFP-2x6His (cellfree expressed))</td><td class="col2"></td>
</tr>
<tr class="row2">
<td class="col0"> 4-6 </td><td class="col1">neg. control (cellfree Expression without DNA)</td><td class="col2"></td>
</tr>
<tr class="row3">
<td class="col0"> 7-8</td><td class="col1"> His-GFP Lysate </td><td class="col2"></td>
</tr>
<tr class="row4">
<td class="col0"> 9 </td><td class="col1">n. T. GFP Lysate) </td><td class="col2"></td>
</tr>
<tr class="row5">
<td class="col0"> 10 </td><td class="col1">bBSA</td><td class="col2 rightalign">  0.2 mg/ml </td>
</tr>
<tr class="row6">
<td class="col0"> 11 </td><td class="col1">Max GFP</td><td class="col2"> 0.5 mg/ml </td>
</tr>
</table></div>
<!-- EDIT231 TABLE [87837-88074] -->
</div>
<!-- EDIT230 SECTION "2015.08.13" [87623-88075] -->
<h3 class="sectionedit232">2015.08.14</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA in PBS for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked with 10 mg/mL BSA in PBS for 30 min</div>
</li>
<li class="level1"><div class="li"> slides were washed in PBS for 10 min</div>
</li>
<li class="level1"><div class="li"> slides were washed in d1/10 diluted PBS with 20 mM Imidazole</div>
</li>
<li class="level1"><div class="li"> slides were washed in d1/10 diluted PBS </div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<div class="table sectionedit233"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">a-GFP</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">StrepCy5</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT233 TABLE [88412-88644] -->
</div>
<h4>Results</h4>
<div class="level4">
</div>
<!-- EDIT232 SECTION "2015.08.14" [88076-88663] -->
<h2 class="sectionedit234">Experiment 46a: Ni-NTA slides for iRIf - cell free expressed tYFP</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT234 SECTION "Experiment 46a: Ni-NTA slides for iRIf - cell free expressed tYFP" [88664-88864] -->
<h3 class="sectionedit235">2015.08.09</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 Ni-NTA slides for measurement of cell free expressed Proteins (GFP/tYFP) or antigen lysate</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 2 PDITC slides</div>
</li>
<li class="level1"><div class="li"> plasma activation: 80 L/h</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 45 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2h</div>
</li>
<li class="level1"><div class="li"> slides were sandwiched with NTA solution, 1 slide was sandwiched with a normal glass slide</div>
</li>
</ul>
</div>
<!-- EDIT235 SECTION "2015.08.09" [88865-89248] -->
<h3 class="sectionedit236">2015.08.10</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> slides were incubated in blocking solution  for 1 h</div>
</li>
<li class="level1"><div class="li"> slides were incubated in Nickel sulfate solution for 1 h</div>
</li>
<li class="level1"><div class="li"> slides were washed 2x in PBS and 1x in diluted PBS</div>
</li>
<li class="level1"><div class="li"> slides were dried and stored under N<sub>2</sub>-atmosphere</div>
</li>
</ul>
<ul>
<li class="level1"><div class="li"> spotted slide 254 and 215 with cell free expressed proteins</div>
</li>
<li class="level1"><div class="li"> 3 µL of cell free expressed His-tYFP-Halo and controls (His-GFP, bBSA) were spotted and incubated o/n</div>
</li>
</ul>
<p>
spotting pattern: 
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" width="300"/></a> <br/>
</p>
<div class="table sectionedit237"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1</td><td class="col1"> His-tYFP-Halo KK </td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0">2</td><td class="col1"> His-tYFP-Halo KK + Mg </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0">3</td><td class="col1"> His-tYFP-Halo BK </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0">4</td><td class="col1"> His-tYFP-Halo BK + Mg </td><td class="col2"> </td>
</tr>
<tr class="row5">
<td class="col0">5</td><td class="col1"> His-tYFP-Halo neg. </td><td class="col2"> </td>
</tr>
<tr class="row6">
<td class="col0">6</td><td class="col1"> bBSA </td><td class="col2"> 200 µg/ml </td>
</tr>
<tr class="row7">
<td class="col0">7</td><td class="col1"> Max His-GFP </td><td class="col2"> 0.5 mg/mL </td>
</tr>
<tr class="row8">
<td class="col0">8</td><td class="col1"> His-GFP Lysate </td><td class="col2"> ~1 mg/mL </td>
</tr>
<tr class="row9">
<td class="col0">9</td><td class="col1"> His-GFP Lysate </td><td class="col2"> ~1 mg/mL </td>
</tr>
<tr class="row10">
<td class="col0">10</td><td class="col1"> Max His-GFP </td><td class="col2"> 1 mg/mL </td>
</tr>
<tr class="row11">
<td class="col0">11</td><td class="col1"> nT-GFP Lysate </td><td class="col2"> ~1 mg/mL </td>
</tr>
</table></div>
<!-- EDIT237 TABLE [89794-90156] -->
</div>
<!-- EDIT236 SECTION "2015.08.10" [89249-90157] -->
<h3 class="sectionedit238">2015.08.11</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> sildes were blocked with 10 mg/mL BSA for 30 min</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS for 10 min</div>
</li>
<li class="level1"><div class="li"> slides were washed with 20 mM Imidazole in diluted PBS for 10 min</div>
</li>
<li class="level1"><div class="li"> slides were washed with diluted PBS for 10 min</div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<div class="table sectionedit239"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">a-tYFP</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">a-GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">StrepCy5</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT239 TABLE [90499-90778] -->
</div>
<h4>Results</h4>
<div class="level4">
</div>
<!-- EDIT238 SECTION "2015.08.11" [90158-90797] -->
<h2 class="sectionedit240">Experiment 45b: Ni-NTA slides for iRIf - cell free tYFP</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT240 SECTION "Experiment 45b: Ni-NTA slides for iRIf - cell free tYFP" [90798-90988] -->
<h3 class="sectionedit241">2015.08.07</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 Ni-NTA slides for measurement of cell free expressed proteins and purified antigens in iRIf</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 2 PDITC slides according to protocol</div>
</li>
<li class="level1"><div class="li"> plasma activation: 80 L/h</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
<li class="level1"><div class="li"> slides were incubated with NTA-solution o/n</div>
</li>
</ul>
</div>
<!-- EDIT241 SECTION "2015.08.07" [90989-91351] -->
<h3 class="sectionedit242">2015.08.08</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocking in APTES blocking solution for 1h at RT</div>
</li>
<li class="level1"><div class="li"> NiSO<sub>4</sub> incubation time: 1 h at RT</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS/diluted PBS</div>
</li>
</ul>
</div>
<!-- EDIT242 SECTION "2015.08.08" [91352-91546] -->
<h3 class="sectionedit243">2015.08.09</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> used slide 424 and 029</div>
</li>
<li class="level1"><div class="li"> 3 µL of cell free expressed His-tYFP-Halo and controls (His-GFP, bBSA) were spotted and incubated o/n</div>
</li>
</ul>
<p>
spotting pattern: 
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" width="300"/></a> <br/>
</p>
<div class="table sectionedit244"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1</td><td class="col1"> His-tYFP-Spy KK </td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0">2</td><td class="col1"> His-tYFP-Spy KK + Mg </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0">3</td><td class="col1"> His-tYFP-Spy BK </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0">4</td><td class="col1"> His-tYFP-Spy BK + Mg </td><td class="col2"> </td>
</tr>
<tr class="row5">
<td class="col0">5</td><td class="col1"> His-tYFP-Spy neg. </td><td class="col2"> </td>
</tr>
<tr class="row6">
<td class="col0">6</td><td class="col1"> bBSA </td><td class="col2"> 200 µg/ml </td>
</tr>
<tr class="row7">
<td class="col0">7</td><td class="col1"> Max His-GFP </td><td class="col2"> 0.5 mg/mL </td>
</tr>
<tr class="row8">
<td class="col0">8</td><td class="col1"> His-GFP Lysate </td><td class="col2"> ~1 mg/mL </td>
</tr>
<tr class="row9">
<td class="col0">9</td><td class="col1"> His-GFP Lysate </td><td class="col2"> ~1 mg/mL </td>
</tr>
<tr class="row10">
<td class="col0">10</td><td class="col1"> Max His-GFP </td><td class="col2"> 1 mg/mL </td>
</tr>
<tr class="row11">
<td class="col0">11</td><td class="col1"> nT-GFP Lysate </td><td class="col2"> ~1 mg/mL </td>
</tr>
</table></div>
<!-- EDIT244 TABLE [91817-92174] -->
</div>
<!-- EDIT243 SECTION "2015.08.09" [91547-92176] -->
<h3 class="sectionedit245">2015.08.10</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked with 10 mg/mL BSA for 30 min</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS, imidazole in diluted PBS and in diluted PBS</div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<div class="table sectionedit246"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">a-tYFP</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">a-GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">StrepCy5</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT246 TABLE [92432-92711] -->
</div>
<h4>Results</h4>
<div class="level4">
</div>
<!-- EDIT245 SECTION "2015.08.10" [92177-92730] -->
<h2 class="sectionedit247">Experiment 45a: Ni-NTA slides for iRIf - cell free GFP</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT247 SECTION "Experiment 45a: Ni-NTA slides for iRIf - cell free GFP" [92731-92920] -->
<h3 class="sectionedit248">2015.08.07</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 Ni-NTA slides for measurement of cell free expressed proteins </div>
</li>
<li class="level1"><div class="li"> slide 024 and 216</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 2 PDITC slides according to protocol</div>
</li>
<li class="level1"><div class="li"> plasma activation: 80 L/h</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
<li class="level1"><div class="li"> slides were incubated with NTA-solution o/n</div>
</li>
</ul>
</div>
<!-- EDIT248 SECTION "2015.08.07" [92921-93276] -->
<h3 class="sectionedit249">2015.08.08</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocking in APTES blocking solution for 1h at RT</div>
</li>
<li class="level1"><div class="li"> NiSO<sub>4</sub> incubation time: 1 h at RT</div>
</li>
<li class="level1"><div class="li"> slides were washed with PBS/diluted PBS</div>
</li>
<li class="level1"><div class="li"> 3 µL of cell free expressed His-GFP and controls (His-GFP, bBSA) were spotted and incubated o/n</div>
</li>
</ul>
<p>
spotting pattern: 
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" width="300"/></a> <br/>
</p>
<div class="table sectionedit250"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1</td><td class="col1"> HA-GFP-His KK </td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0">2</td><td class="col1"> HA-GFP-His KK+Mg </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0">3</td><td class="col1"> HA-GFP-His BK </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0">4</td><td class="col1"> HA-GFP-His BK+Mg </td><td class="col2"> </td>
</tr>
<tr class="row5">
<td class="col0">5</td><td class="col1"> HA-GFP-His BP </td><td class="col2"> </td>
</tr>
<tr class="row6">
<td class="col0">6</td><td class="col1"> HA-GFP-His PP </td><td class="col2"> </td>
</tr>
<tr class="row7">
<td class="col0">7</td><td class="col1"> Max His-GFP </td><td class="col2"> 0.1 mg/mL </td>
</tr>
<tr class="row8">
<td class="col0">8</td><td class="col1"> nT-GFP Lysate </td><td class="col2"> ~1 mg/mL </td>
</tr>
<tr class="row9">
<td class="col0">9</td><td class="col1"> His-GFP Lysate </td><td class="col2"> ~1 mg/mL </td>
</tr>
<tr class="row10">
<td class="col0">10</td><td class="col1"> cell free neg. </td><td class="col2"> </td>
</tr>
<tr class="row11">
<td class="col0">11</td><td class="col1"> bBSA </td><td class="col2"> 100 µg/ml </td>
</tr>
</table></div>
<!-- EDIT250 TABLE [93659-93968] -->
</div>
<!-- EDIT249 SECTION "2015.08.08" [93277-93969] -->
<h3 class="sectionedit251">2015.08.09</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x 5 min with 10 mg/mL BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked 10 mg/mL BSA for 30 min</div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<div class="table sectionedit252"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">a-GFP</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">StrepCy5</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT252 TABLE [94149-94381] -->
</div>
<h4>Results</h4>
<div class="level4">
</div>
<!-- EDIT251 SECTION "2015.08.09" [93970-94399] -->
<h2 class="sectionedit253">Experiment 44c: PDITC slides for iRIf - own setup 2</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT253 SECTION "Experiment 44c: PDITC slides for iRIf - own setup 2" [94400-94586] -->
<h3 class="sectionedit254">2015.08.07</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 PDITC slides for measurement of purified antigens and PhyA in iRIf and bBSA in own setup</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 2 PDITC slides according to protocol</div>
</li>
<li class="level1"><div class="li"> plasma activation: 80 L/h</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
<li class="level1"><div class="li"> slides were stored in desiccator o/n</div>
</li>
</ul>
</div>
<!-- EDIT254 SECTION "2015.08.07" [94587-94939] -->
<h3 class="sectionedit255">2015.08.09</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spotted the slides with 200 µg/mL bBSA</div>
</li>
<li class="level1"><div class="li"> 9 spots with bBSA, 1 spot with BSA (10 mg/mL) in the middle of the slide</div>
</li>
<li class="level1"><div class="li"> slides were incubated at 4 °C o/n</div>
</li>
</ul>
</div>
<!-- EDIT255 SECTION "2015.08.09" [94940-95149] -->
<h3 class="sectionedit256">2015.08.10</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocked spots 2x with 10 mg/mL BSA for 20 min</div>
</li>
<li class="level1"><div class="li"> blocked slides with 10 mg/mL BSA for 45 min </div>
</li>
<li class="level1"><div class="li"> dried slides with wafergun and stored them under N<sub>2</sub>-atmosphere till measurement</div>
</li>
</ul>
</div>
<!-- EDIT256 SECTION "2015.08.10" [95150-95393] -->
<h2 class="sectionedit257">Experiment 44b: PDITC slides for iRIf - antigens</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT257 SECTION "Experiment 44b: PDITC slides for iRIf - antigens" [95394-95577] -->
<h3 class="sectionedit258">2015.08.07</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 PDITC slides for measurement of purified antigens and PhyA in iRIf</div>
</li>
<li class="level1"><div class="li"> slide 265 and 426</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 2 PDITC slides according to protocol</div>
</li>
<li class="level1"><div class="li"> plasma activation: 80 L/h</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
<li class="level1"><div class="li"> slides were stored in desiccator o/n</div>
</li>
</ul>
</div>
<!-- EDIT258 SECTION "2015.08.07" [95578-95930] -->
<h3 class="sectionedit259">2015.08.08</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> antigens were diluted to a concentration of ~0.4-0.5 mg/mL</div>
</li>
<li class="level1"><div class="li"> 3µL of antigens and controls (GFP, bBSA) were spotted and icubated o/n</div>
</li>
<li class="level1"><div class="li"> spinned antigens before spotting</div>
</li>
</ul>
<p>
Spotting pattern:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="/igem2015/lib/exe/fetch.php?w=400&amp;media=labjournal:surchem:surchem_11_slide_usage.png" width="400"/></a>
</p>
<div class="table sectionedit260"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> Elution no. </th><th class="col3"> Concentration </th>
</tr>
<tr class="row1">
<td class="col0"> 1-2 </td><td class="col1"> HIV(17) </td><td class="col2 leftalign"> </td><td class="col3"> 0.4-0.5 mg/ml </td>
</tr>
<tr class="row2">
<td class="col0"> 3-4 </td><td class="col1"> HCV(10) </td><td class="col2 leftalign"> </td><td class="col3"> 0.4-0.5 mg/ml </td>
</tr>
<tr class="row3">
<td class="col0"> 5-6 </td><td class="col1"> Tetanus(11) </td><td class="col2 leftalign"> </td><td class="col3"> 0.4-0.5 mg/ml </td>
</tr>
<tr class="row4">
<td class="col0"> 7 </td><td class="col1"> GFP </td><td class="col2 leftalign"> </td><td class="col3"> 0.1 mg/ml </td>
</tr>
<tr class="row5">
<td class="col0"> 8 </td><td class="col1"> PhyA </td><td class="col2 leftalign"> </td><td class="col3"> 0.4-0.5 mg/ml </td>
</tr>
<tr class="row6">
<td class="col0"> 9 </td><td class="col1"> bBSA </td><td class="col2 leftalign"> </td><td class="col3"> 0.1 mg/ml </td>
</tr>
<tr class="row7">
<td class="col0"> 10-11 </td><td class="col1"> Salmonella(15) </td><td class="col2"> </td><td class="col3"> 0.4-0.5 mg/ml </td>
</tr>
</table></div>
<!-- EDIT260 TABLE [96239-96532] -->
</div>
<!-- EDIT259 SECTION "2015.08.08" [95931-96533] -->
<h3 class="sectionedit261">2015.08.09</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<p>
Slide 265 - measured in new setup
</p>
<div class="table sectionedit262"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">720</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">Anti-HIV (rabbit,pk))</td><td class="col1">4</td><td class="col2">45</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">45</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">Anti-HCV (rabbit,pk)</td><td class="col1">6</td><td class="col2">45</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">45</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">Anti-phyA (rabbit,N-term.)</td><td class="col1">8</td><td class="col2">45</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">45</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row10">
<td class="col0">Anti-Rabbit</td><td class="col1">10</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row11">
<td class="col0">Buffer</td><td class="col1">11</td><td class="col2">45</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row12">
<td class="col0">Anti-HIV (mouse, monoklonal)</td><td class="col1">12</td><td class="col2">45</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row13">
<td class="col0">Buffer</td><td class="col1">13</td><td class="col2">45</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row14">
<td class="col0">Anti-HCV (mouse, monoklonal)</td><td class="col1">14</td><td class="col2">45</td><td class="col3">600</td><td class="col4">10 ug/ml</td>
</tr>
<tr class="row15">
<td class="col0">Buffer</td><td class="col1">15</td><td class="col2">45</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row16">
<td class="col0">Anti-Tetanus (mouse, monoklonal)</td><td class="col1">16</td><td class="col2">45</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row17">
<td class="col0">Buffer</td><td class="col1">17</td><td class="col2">45</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row18">
<td class="col0">Anti-GFP (goat)</td><td class="col1">18</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row19">
<td class="col0">Buffer</td><td class="col1">19</td><td class="col2">45</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row20">
<td class="col0">StrepCy</td><td class="col1">20</td><td class="col2">45</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row21">
<td class="col0">Buffer</td><td class="col1">21</td><td class="col2">45</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row22">
<td class="col0">Anti-Mouse</td><td class="col1">22</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row23">
<td class="col0">Buffer</td><td class="col1">23</td><td class="col2">45</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT262 TABLE [96617-97382] -->
</div>
<!-- EDIT261 SECTION "2015.08.09" [96534-97382] -->
<h2 class="sectionedit263">Experiment 44a: PDITC slides for iRIf - own setup 1</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<!-- EDIT263 SECTION "Experiment 44a: PDITC slides for iRIf - own setup 1" [97383-97569] -->
<h3 class="sectionedit264">2015.08.07</h3>
<div class="level3">
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 PDITC slides for measurement of purified antigens and PhyA in iRIf and bBSA in own setup</div>
</li>
</ul>
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepared 2 PDITC slides according to protocol</div>
</li>
<li class="level1"><div class="li"> plasma activation: 80 L/h</div>
</li>
<li class="level1"><div class="li"> APTES incubation: 30 min</div>
</li>
<li class="level1"><div class="li"> PDITC incubation: 2 h</div>
</li>
<li class="level1"><div class="li"> slides were stored in desiccator o/n</div>
</li>
</ul>
</div>
<!-- EDIT264 SECTION "2015.08.07" [97570-97922] -->
<h3 class="sectionedit265">2015.08.08</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spotted the slides with 200 µg/mL bBSA</div>
</li>
<li class="level1"><div class="li"> 9 spots with bBSA, 1 spot with BSA (10 mg/mL) in the middle of the slide</div>
</li>
<li class="level1"><div class="li"> slides were incubated at 4 °C for 11 h</div>
</li>
<li class="level1"><div class="li"> slides were blocked and afterwards measured in own setup</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
</div>
<!-- EDIT265 SECTION "2015.08.08" [97923-98215] -->
<h2 class="sectionedit266">Experiment 43: Ni/NTA slides for Normann</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 2 iRIf slides from Normann (309, 310) </div>
</li>
</ul>
</div>
<!-- EDIT266 SECTION "Experiment 43: Ni/NTA slides for Normann" [98216-98466] -->
<h3 class="sectionedit267">2015.08.04</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 6 iRIf slides were cleaned according to <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Slide_Wash" title="protocols:slide_wash">iRIf slide wash steps</a></div>
</li>
<li class="level1"><div class="li"> plasma activation for 5 min at 80 L/h gasflow</div>
</li>
<li class="level1"><div class="li"> incubated in APTES solution for 30 min </div>
</li>
<li class="level1"><div class="li"> incubation in PDITC solution for 2.5 h</div>
</li>
<li class="level1"><div class="li"> sandwiched slides with 80 µL NTA and incubated them at 4°C o/n </div>
</li>
</ul>
</div>
<!-- EDIT267 SECTION "2015.08.04" [98467-98812] -->
<h3 class="sectionedit268">2015.08.05</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> blocked slides for 1 h in APTES blocking solution</div>
</li>
<li class="level1"><div class="li"> washed 2x in PBS for 10 min</div>
</li>
<li class="level1"><div class="li"> incubated in NiSO<sub>4</sub>-solution for 1 h</div>
</li>
<li class="level1"><div class="li"> washed 1x in PBS for 10 min</div>
</li>
<li class="level1"><div class="li"> washed 2x in diluted PBS for 10 min then dried with wafer gun</div>
</li>
<li class="level1"><div class="li"> stored slides in slideholder under N<sub>2</sub>-atmosphere for Norman (uses them the next day)</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
</div>
<!-- EDIT268 SECTION "2015.08.05" [98813-99214] -->
<h2 class="sectionedit269">Experiment 42c: washed iRIf slides for Halo-surface (Piehler) with Günter</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Slide_Wash" title="protocols:slide_wash">iRIf slide wash steps</a></div>
</li>
</ul>
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> wash six of <strong>Jürgens iRIf slides</strong> (254, 466, 423, 426, 253 ,424) for first test with Halo </div>
</li>
</ul>
</div>
<!-- EDIT269 SECTION "Experiment 42c: washed iRIf slides for Halo-surface (Piehler) with Günter" [99215-99491] -->
<h3 class="sectionedit270">2015.08.05</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 2 slides: 423, 466</div>
</li>
<li class="level1"><div class="li"> slides were cleaned with acetone, isopropanol and aqua dest </div>
</li>
<li class="level1"><div class="li"> plasma activation for 10 min at gas flow 60 L/h with valve 2 (air)</div>
</li>
<li class="level1"><div class="li"> slides were cross-sandwiched (Piehler style) with 15 µL of PLL-PEG-HTL for 15 min</div>
</li>
<li class="level1"><div class="li"> 3 µL of Halo-GFP-His, Halo-mCherry-His, bBSA, and 2 µL of pos./neg. controls (from Piehler: His-Halo-GFP/His-GFP) were spotted</div>
</li>
</ul>
<p>
spotting pattern: 
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" width="300"/></a> <br/>
</p>
<div class="table sectionedit271"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-2</td><td class="col1"> Halo-GFP 1:20 </td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0">3-4</td><td class="col1"> Halo-GFP 1:10 </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0">5-6</td><td class="col1"> Halo-mCHERRY 1:20 </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0">7</td><td class="col1"> His-Halo-GFP Piehler </td><td class="col2"> </td>
</tr>
<tr class="row5">
<td class="col0">8</td><td class="col1"> His-GFP Piehler </td><td class="col2"> </td>
</tr>
<tr class="row6">
<td class="col0">9</td><td class="col1"> bBSA </td><td class="col2"> 100 µg/ml </td>
</tr>
<tr class="row7">
<td class="col0">10-11</td><td class="col1"> Halo-mCHERRY 1:10 </td><td class="col2"> </td>
</tr>
</table></div>
<!-- EDIT271 TABLE [100007-100219] --><ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked in 10 mg/ml BSA for 30 min</div>
</li>
<li class="level1"><div class="li"> slide were washed with ddH<sub>2</sub>O</div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
Slide 423 - measured in new setup
</p>
<div class="table sectionedit272"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">a-His (from AG Roth)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">a-GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">StrepCy</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT272 TABLE [100422-100714] -->
<p>
Slide 466 - measured in old setup
</p>
<div class="table sectionedit273"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">a-His (from iGEM Lab)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">a-GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">StrepCy</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT273 TABLE [100750-101043] --><ul>
<li class="level1"><div class="li"> Cy5 Fluorescence was measured afterwards</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/7a/Freiburg_labjournal-surchem-exp_42c_423.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42c_423.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 423: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/7d/Freiburg_labjournal-surchem-exp_42c_423_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42c_423_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 423: Picture obtained from fluorescence scanner. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/d/da/Freiburg_labjournal-surchem-exp_42c_423_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42c_423_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 423: Binding curves</div></div></div><hr/>
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/8/83/Freiburg_labjournal-surchem-exp_42c_466.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42c_466.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 466: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/9/9a/Freiburg_labjournal-surchem-exp_42c_466_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42c_466_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 466: Picture obtained from fluorescence scanner. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/45/Freiburg_labjournal-surchem-exp_42c_466_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42c_466_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 466: Binding curves</div></div></div>
<p>
→ plasmaactivation with pure air does not lead to better binding of PLL-PEG-HTL to the surface
</p>
</div>
<!-- EDIT270 SECTION "2015.08.05" [99492-101883] -->
<h2 class="sectionedit274">Experiment 42b: washed iRIf slides for Halo-surface (Piehler) with Günter</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Slide_Wash" title="protocols:slide_wash">iRIf slide wash steps</a></div>
</li>
</ul>
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> wash six of <strong>Jürgens iRIf slides</strong> (254, 466, 423, 426, 253 ,424) for first test with Halo </div>
</li>
</ul>
</div>
<!-- EDIT274 SECTION "Experiment 42b: washed iRIf slides for Halo-surface (Piehler) with Günter" [101884-102160] -->
<h3 class="sectionedit275">2015.08.05</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 2 slides: 426, 424</div>
</li>
<li class="level1"><div class="li"> slides were cleaned with acetone, isopropanol and aqua dest </div>
</li>
<li class="level1"><div class="li"> plasma activation for 10 min at gas flow 60 L/h with valve 1 (steam)  </div>
</li>
<li class="level1"><div class="li"> slides were cross-sandwiched (Piehler style) with 15 µL of PLL-PEG-HTL for 15 min</div>
</li>
<li class="level1"><div class="li"> 3 µL of Halo-GFP-His, Halo-mCherry-His, bBSA, and 2 µL of pos./neg. controls (from Piehler: His-Halo-GFP/His-GFP) were spotted</div>
</li>
</ul>
<p>
spotting pattern: 
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" width="300"/></a> <br/>
</p>
<div class="table sectionedit276"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-2</td><td class="col1"> Halo-GFP 1:20 </td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0">3-4</td><td class="col1"> Halo-GFP 1:10 </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0">5-6</td><td class="col1"> Halo-mCHERRY 1:20 </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0">7</td><td class="col1"> His-Halo-GFP Piehler </td><td class="col2"> </td>
</tr>
<tr class="row5">
<td class="col0">8</td><td class="col1"> His-GFP Piehler </td><td class="col2"> </td>
</tr>
<tr class="row6">
<td class="col0">9</td><td class="col1"> bBSA </td><td class="col2"> 100 µg/ml </td>
</tr>
<tr class="row7">
<td class="col0">10-11</td><td class="col1"> Halo-mCHERRY 1:10 </td><td class="col2"> </td>
</tr>
</table></div>
<!-- EDIT276 TABLE [102680-102892] -->
<p>
slide 426
</p>
<ul>
<li class="level1"><div class="li"> slide wasn't blocked at all → measured directly</div>
</li>
</ul>
<p>
slide 424
</p>
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL for 5 min</div>
</li>
<li class="level1"><div class="li"> slide was blocked in 10 mg/ml BSA for 30 min</div>
</li>
<li class="level1"><div class="li"> slide was washed with ddH<sub>2</sub>O</div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
Slide 426 - measured in old setup
</p>
<ul>
<li class="level1"><div class="li"> slide wasn't blocked before measurement</div>
</li>
</ul>
<div class="table sectionedit277"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">a-GFP</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">StrepCy5</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT277 TABLE [103209-103441] -->
<p>
Slide 424  - measured in old setup
(Strep-Cy5 and a-GFP step were accidently switched)
</p>
<div class="table sectionedit278"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">a-His</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">StrepCy</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">a-GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT278 TABLE [103531-103808] --><ul>
<li class="level1"><div class="li"> Cy5 Fluorescence was measured afterwards</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/c/c8/Freiburg_labjournal-surchem-exp_42b_slide424.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42b_slide424.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 424: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/b/b5/Freiburg_labjournal-surchem-exp_42b_slide424_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42b_slide424_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 424: Picture obtained from fluorescence scanner. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/d/da/Freiburg_labjournal-surchem-exp_42b_slide424_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42b_slide424_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 424: Binding curves</div></div></div><hr/>
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/1/17/Freiburg_labjournal-surchem-exp_42b_slide426.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42b_slide426.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 426: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/1/10/Freiburg_labjournal-surchem-exp_42b_slide426_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42b_slide426_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 426: Picture obtained from fluorescence scanner. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/7b/Freiburg_labjournal-surchem-exp_42b_slide426_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42b_slide426_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 426: Binding curves</div></div></div><ul>
<li class="level1"><div class="li"> for slide 426: during the BSA step lots of BSA bound to the surface –&gt; should block before</div>
</li>
</ul>
</div>
<!-- EDIT275 SECTION "2015.08.05" [102161-104678] -->
<h2 class="sectionedit279">Experiment 42a: washed iRIf slides for Halo-surface (Silan by abcr) with Günter</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Slide_Wash" title="protocols:slide_wash">iRIf slide wash steps</a></div>
</li>
</ul>
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> wash six of <strong>Jürgens iRIf slides</strong> (254, 466, 423, 426, 253 ,424) for first test with Halo </div>
</li>
</ul>
</div>
<!-- EDIT279 SECTION "Experiment 42a: washed iRIf slides for Halo-surface (Silan by abcr) with Günter" [104679-104961] -->
<h3 class="sectionedit280">2015.08.05</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 2 slides: 254, 253</div>
</li>
<li class="level1"><div class="li"> slides were cleaned with acetone, isopropanol and aqua dest </div>
</li>
<li class="level1"><div class="li"> plasma activation for 10 min at gas flow 60 L/h with valve 1 (steam)</div>
</li>
<li class="level1"><div class="li"> slides were sandwiched with 80 µL of abcr-silane and incubated for 1 h</div>
</li>
<li class="level1"><div class="li"> washed 30s in acetone</div>
</li>
<li class="level1"><div class="li"> dried with N<sub>2</sub></div>
</li>
<li class="level1"><div class="li"> 3 µl of Halo-GFP, Halo-mCHERRY and bBSA as well as 2 µl of His-Halo-GFP (Piehler positive control) and His-GFP (Piehler negative control) were spotted</div>
</li>
</ul>
<p>
spotting pattern: 
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" width="300"/></a> <br/>
</p>
<div class="table sectionedit281"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-2</td><td class="col1"> Halo-GFP 1:20 </td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0">3-4</td><td class="col1"> Halo-GFP 1:10 </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0">5-6</td><td class="col1"> Halo-mCHERRY 1:20 </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0">7</td><td class="col1"> His-Halo-GFP Piehler </td><td class="col2"> </td>
</tr>
<tr class="row5">
<td class="col0">8</td><td class="col1"> His-GFP Piehler </td><td class="col2"> </td>
</tr>
<tr class="row6">
<td class="col0">9</td><td class="col1"> bBSA </td><td class="col2"> 100 µg/ml </td>
</tr>
<tr class="row7">
<td class="col0">10-11</td><td class="col1"> Halo-mCHERRY 1:10 </td><td class="col2"> </td>
</tr>
</table></div>
<!-- EDIT281 TABLE [105546-105758] --><ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked in 10 mg/ml BSA for 30 min</div>
</li>
<li class="level1"><div class="li"> slide were washed with ddH<sub>2</sub>O</div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
slide 254 - measured with new setup
</p>
<div class="table sectionedit282"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">a-His (from AG Roth)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">a-GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">StrepCy</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT282 TABLE [105963-106255] -->
<p>
slide 253 - measured with new setup
</p>
<div class="table sectionedit283"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">a-His (from AG Roth)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">a-GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">StrepCy</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT283 TABLE [106294-106586] --><ul>
<li class="level1"><div class="li"> Cy5 Fluorescence was measured afterwards</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/4a/Freiburg_labjournal-surchem-exp_42a_slide253.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42a_slide253.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 253: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/a/ad/Freiburg_labjournal-surchem-exp_42a_slide253_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42a_slide253_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 253: Picture obtained from fluorescence scanner. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/c/c2/Freiburg_labjournal-surchem-exp_42a_slide253_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42a_slide253_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 253: Binding curves</div></div></div><hr/>
<div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/7d/Freiburg_labjournal-surchem-exp_42a_slide254.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42a_slide254.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 253: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/8/80/Freiburg_labjournal-surchem-exp_42a_slide254_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42a_slide254_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 253: Picture obtained from fluorescence scanner. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><div class="thumb2 trien" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/e/e4/Freiburg_labjournal-surchem-exp_42a_slide254_binding.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_42a_slide254_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 253: Binding curves</div></div></div>
</div>
<!-- EDIT280 SECTION "2015.08.05" [104962-107358] -->
<h2 class="sectionedit284">Experiment 41b: PDITC iRIf slides for Halo-surface (Wiesmüller Halo 2) with Günter</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 6 of <strong>our iRIf slides</strong> with APTES/PDITC surface for first test with Halo</div>
</li>
</ul>
</div>
<!-- EDIT284 SECTION "Experiment 41b: PDITC iRIf slides for Halo-surface (Wiesmüller Halo 2) with Günter" [107359-107689] -->
<h3 class="sectionedit285">2015.08.04</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 6 iRIf slides were cleaned according to <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Slide_Wash" title="protocols:slide_wash">iRIf slide wash steps</a></div>
</li>
<li class="level1"><div class="li"> plasma activation for 5 min at 80 L/h gasflow</div>
</li>
<li class="level1"><div class="li"> incubated in APTES solution for 40 min</div>
</li>
<li class="level1"><div class="li"> incubation in PDITC solution for 2.5 h</div>
</li>
<li class="level1"><div class="li"> slides were stored in desiccator o/n</div>
</li>
</ul>
</div>
<!-- EDIT285 SECTION "2015.08.04" [107690-108005] -->
<h3 class="sectionedit286">2015.08.05</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 3 slides: 287, 215, 315</div>
</li>
<li class="level1"><div class="li"> slides were incubated in slideholder with Halo 2-solution for 2 days</div>
</li>
</ul>
</div>
<!-- EDIT286 SECTION "2015.08.05" [108006-108156] -->
<h3 class="sectionedit287">2015.08.07</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 3 µL of cell free expressed proteins, Halo-GFP, Halo-mCherry and pos/neg control (from Piehler) were spotted on slides 215 and 315</div>
</li>
<li class="level1"><div class="li"> slides were incubated for 1 h at RT</div>
</li>
</ul>
<p>
spotting pattern: 
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" width="300"/></a> <br/>
</p>
<div class="table sectionedit288"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-2</td><td class="col1"> Koko pos. </td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0">3-4</td><td class="col1"> Koko neg. </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0">5-6</td><td class="col1"> Promega pos. </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0">7</td><td class="col1"> Halo-GFP (pos, Piehler) </td><td class="col2"> </td>
</tr>
<tr class="row5">
<td class="col0">8</td><td class="col1"> His-GFP (neg, Piehler) </td><td class="col2"> </td>
</tr>
<tr class="row6">
<td class="col0">9</td><td class="col1"> bBSA </td><td class="col2"> 100 µg/ml </td>
</tr>
<tr class="row7">
<td class="col0">10</td><td class="col1"> Halo-GFP 1:10 </td><td class="col2"> </td>
</tr>
<tr class="row8">
<td class="col0">11</td><td class="col1"> Halo-mCHERRY 1:10 </td><td class="col2"> </td>
</tr>
</table></div>
<!-- EDIT288 TABLE [108469-108698] --><ul>
<li class="level1"><div class="li"> slide 287 was first blocked with 10 mg/mL BSA for 30 min</div>
</li>
<li class="level1"><div class="li"> then 3 µL of Halo-GFP, His-GFP, pos/neg control (from Piehler), His-GFP and mCherry were spotted on slide 287</div>
</li>
<li class="level1"><div class="li"> slide was incubated for 1 h at RT</div>
</li>
</ul>
<p>
spotting pattern: 
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" width="300"/></a> <br/>
</p>
<div class="table sectionedit289"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-3</td><td class="col1"> Halo-GFP </td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0">4-6</td><td class="col1"> Halo-mCherry </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0">7</td><td class="col1"> Halo-GFP (pos, Piehler) </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0">8</td><td class="col1"> His-GFP (neg, Piehler) </td><td class="col2"> </td>
</tr>
<tr class="row5">
<td class="col0">9</td><td class="col1"> bBSA </td><td class="col2"> 100 µg/ml </td>
</tr>
<tr class="row6">
<td class="col0">10</td><td class="col1"> His-GFP </td><td class="col2"> </td>
</tr>
<tr class="row7">
<td class="col0">11</td><td class="col1"> His-mCHERRY </td><td class="col2"> </td>
</tr>
</table></div>
<!-- EDIT289 TABLE [109001-109197] -->
<p>
for all 3 slides:
</p>
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked with 10 mg/mL BSA for 30 min</div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
slide 215 and 315
</p>
<p>
slide 215 was measured in old setup, slide 315 was measured in new setup
</p>
<div class="table sectionedit290"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">a-tYFP</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 µg/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">a-GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 µg/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">StrepCy</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 µg/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT290 TABLE [109439-109720] -->
<p>
slide 287 was measured with the following flush protocol in the new setup
</p>
<div class="table sectionedit291"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (µl/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">a-GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 µg/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">StrepCy</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 µg/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT291 TABLE [109796-110030] --><ul>
<li class="level1"><div class="li"> Cy5 Fluorescence was measured afterwards</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/c/c4/Freiburg_labjournal-surchem-exp_41_b_215.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_41_b_215.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 215: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/e/ef/Freiburg_labjournal-surchem-exp_41_b_215_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_41_b_215_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 215: Picture obtained from fluorescence scanner. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/3/3d/Freiburg_labjournal-surchem-exp_41_b_287.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_41_b_287.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 287: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/0/0f/Freiburg_labjournal-surchem-exp_41_b_287_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_41_b_287_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 287: Picture obtained from fluorescence scanner. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/23/Freiburg_labjournal-surchem-exp_41_b_315.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_41_b_315.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 315: Selection of ROI</div></div></div>
</div>
<!-- EDIT287 SECTION "2015.08.07" [108157-110681] -->
<h2 class="sectionedit292">Experiment 41a: PDITC iRIf slides for Halo-surface (Wiesmüller Halo 1) with Günter</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 6 of <strong>our iRIf slides</strong> with APTES/PDITC surface for first test with Halo</div>
</li>
</ul>
</div>
<!-- EDIT292 SECTION "Experiment 41a: PDITC iRIf slides for Halo-surface (Wiesmüller Halo 1) with Günter" [110682-111013] -->
<h3 class="sectionedit293">2015.08.04</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 6 iRIf slides were cleaned according to <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Slide_Wash" title="protocols:slide_wash">iRIf slide wash steps</a></div>
</li>
<li class="level1"><div class="li"> plasma activation for 5 min at 80 L/h gasflow</div>
</li>
<li class="level1"><div class="li"> incubated in APTES solution for 40 min</div>
</li>
<li class="level1"><div class="li"> incubation in PDITC solution for 2.5 h</div>
</li>
<li class="level1"><div class="li"> slides were stored in desiccator o/n</div>
</li>
</ul>
</div>
<!-- EDIT293 SECTION "2015.08.04" [111014-111329] -->
<h3 class="sectionedit294">2015.08.05</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 3 slides: 012, 208, 121</div>
</li>
<li class="level1"><div class="li"> slides were incubated in slideholder with Halo 1-solution for 2 days</div>
</li>
</ul>
</div>
<!-- EDIT294 SECTION "2015.08.05" [111330-111480] -->
<h3 class="sectionedit295">2015.08.07</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 3 µL of cell free expressed proteins, Halo-GFP, Halo-mCherry and pos/neg control (from Piehler) were spotted on slides 208 and 121</div>
</li>
<li class="level1"><div class="li"> slides were incubated for 1 h at RT</div>
</li>
</ul>
<p>
spotting pattern: 
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" width="300"/></a> <br/>
</p>
<div class="table sectionedit296"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-2</td><td class="col1"> Koko pos. </td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0">3-4</td><td class="col1"> Koko neg. </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0">5-6</td><td class="col1"> Promega pos. </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0">7</td><td class="col1"> Halo-GFP (pos, Piehler) </td><td class="col2"> </td>
</tr>
<tr class="row5">
<td class="col0">8</td><td class="col1"> His-GFP (neg, Piehler) </td><td class="col2"> </td>
</tr>
<tr class="row6">
<td class="col0">9</td><td class="col1"> bBSA </td><td class="col2"> 100 µg/ml </td>
</tr>
<tr class="row7">
<td class="col0">10</td><td class="col1"> Halo-GFP 1:10 </td><td class="col2"> </td>
</tr>
<tr class="row8">
<td class="col0">11</td><td class="col1"> Halo-mCHERRY 1:10 </td><td class="col2"> </td>
</tr>
</table></div>
<!-- EDIT296 TABLE [111793-112022] --><ul>
<li class="level1"><div class="li"> slide 012 was first blocked with 10 mg/mL BSA for 30 min</div>
</li>
<li class="level1"><div class="li"> then 3 µL of Halo-GFP, His-GFP, pos/neg control (from Piehler), His-GFP and mCherry were spotted on slide 012</div>
</li>
<li class="level1"><div class="li"> slide was incubated for 1 h at RT</div>
</li>
</ul>
<p>
spotting pattern: 
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" width="300"/></a> <br/>
</p>
<div class="table sectionedit297"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th><th class="col2"> concentration </th>
</tr>
<tr class="row1">
<td class="col0">1-3</td><td class="col1"> Halo-GFP </td><td class="col2"> </td>
</tr>
<tr class="row2">
<td class="col0">4-6</td><td class="col1"> Halo-mCherry </td><td class="col2"> </td>
</tr>
<tr class="row3">
<td class="col0">7</td><td class="col1"> Halo-GFP (pos, Piehler) </td><td class="col2"> </td>
</tr>
<tr class="row4">
<td class="col0">8</td><td class="col1"> His-GFP (neg, Piehler) </td><td class="col2"> </td>
</tr>
<tr class="row5">
<td class="col0">9</td><td class="col1"> bBSA </td><td class="col2"> 100 µg/ml </td>
</tr>
<tr class="row6">
<td class="col0">10</td><td class="col1"> His-GFP </td><td class="col2"> </td>
</tr>
<tr class="row7">
<td class="col0">11</td><td class="col1"> His-mCHERRY </td><td class="col2"> </td>
</tr>
</table></div>
<!-- EDIT297 TABLE [112325-112521] -->
<p>
for all 3 slides:
</p>
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> slides were blocked with 10 mg/mL BSA for 30 min</div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
slide 208 and 121
slide 208 was measured in old setup, slide 121 was measured in new setup
</p>
<div class="table sectionedit298"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">a-tYFP</td><td class="col1">4</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 µg/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">a-GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 µg/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">StrepCy</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 µg/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT298 TABLE [112762-113043] -->
<p>
slide 012 was measured with the following flush protocol in the new setup
</p>
<div class="table sectionedit299"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">60</td><td class="col3">600</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">a-GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">3 µg/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">StrepCy</td><td class="col1">8</td><td class="col2">30</td><td class="col3">600</td><td class="col4">5 µg/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT299 TABLE [113119-113352] --><ul>
<li class="level1"><div class="li"> Cy5 Fluorescence was measured afterwards</div>
</li>
</ul>
</div>
<h4>Results</h4>
<div class="level4">
<div class="thumb2 tleft" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/a/af/Freiburg_labjournal-surchem-exp_41a_121.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_41a_121.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 121: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/d/da/Freiburg_labjournal-surchem-exp_41a_121_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_41a_121_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 121: Picture obtained from fluorescence scanner. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><div class="thumb2 tleft" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/22/Freiburg_labjournal-surchem-exp_41a_208.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_41a_208.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 208: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/4b/Freiburg_labjournal-surchem-exp_41a_208_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_41a_208_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 208: Picture obtained from fluorescence scanner. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div><div class="thumb2 tleft" style="width:410px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/c/ce/Freiburg_labjournal-surchem-exp_41a_012.png" width="400"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_41a_012.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 012: Selection of ROI</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/48/Freiburg_labjournal-surchem-exp_41a_012_scan.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aaugust&amp;media=labjournal:surchem:exp_41a_012_scan.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 012: Picture obtained from fluorescence scanner. Top: Emission with 635 nm; Bottom Emission with 523 nm</div></div></div>
<p>
→ Halo surface is not homogenous enough
</p>
<p>
→ strong unspecific binding occuring 
</p>
</div>
<!-- EDIT295 SECTION "2015.08.07" [111481-114254] -->
<h2 class="sectionedit300">Experiment 40b: PDITC iRIf slides for measurement in own setup</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">iRIf slide preparation</a></div>
</li>
</ul>
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 4 iRIf PDTIC slides for measurement</div>
</li>
<li class="level1"><div class="li"> store slides for future measurements</div>
</li>
<li class="level1"><div class="li"> slides were used for measurement in own setup</div>
</li>
</ul>
</div>
<!-- EDIT300 SECTION "Experiment 40b: PDITC iRIf slides for measurement in own setup" [114255-114615] -->
<h3 class="sectionedit301">2015.08.03</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 4 PDITC iRIf slides were prepared according to protocol</div>
</li>
<li class="level1"><div class="li"> plasma activation for 5 min at 80 L/h</div>
</li>
<li class="level1"><div class="li"> slides were incubated in APTES for 45 min</div>
</li>
<li class="level1"><div class="li"> slides were incubated in PDITC for 2 h, washed with Ethanol and Acetone and stored in the desiccator for 40 min</div>
</li>
<li class="level1"><div class="li"> slides were stored at 4 °C under N<sub>2</sub>-atmosphere</div>
</li>
</ul>
</div>
<!-- EDIT301 SECTION "2015.08.03" [114616-114990] -->
<h3 class="sectionedit302">2015.08.04</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 3 µL of bBSA and BSA were spotted on the slides for measurement in own setup</div>
</li>
</ul>
<p>
spotting pattern:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" width="300"/></a> <br/>
</p>
<div class="table sectionedit303"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot </th>
</tr>
<tr class="row1">
<td class="col0"> 4+5 </td><td class="col1"> bBSA 500 µg/mL </td>
</tr>
<tr class="row2">
<td class="col0"> 6+10 </td><td class="col1"> bBSA 200 µg/mL </td>
</tr>
<tr class="row3">
<td class="col0"> 11 </td><td class="col1"> BSA 10 mg/mL </td>
</tr>
</table></div>
<!-- EDIT303 TABLE [115205-115292] --><ul>
<li class="level1"><div class="li"> slides were incubated at 4 °C o/n</div>
</li>
</ul>
</div>
<!-- EDIT302 SECTION "2015.08.04" [114991-115334] -->
<h2 class="sectionedit304">Experiment 40a: PDITC iRIf slides</h2>
<div class="level2">
<ul>
<li class="level1"><div class="li"> protocols used :<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/Plasmaactivation" title="protocols:plasma_activation">Plasma activation</a>, <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Protocols/SlidePrep" title="irif_slide_preparation">[[protocols:irif_slide_preparation|iRIf slide preparation]</a>]</div>
</li>
</ul>
</div>
<h4>Considerations</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> prepare 4 iRIf PDTIC slides for measurement</div>
</li>
<li class="level1"><div class="li"> spot antibodies that shall be detected with antigens (opposite to Exp 37a) on 2 slides</div>
</li>
</ul>
</div>
<!-- EDIT304 SECTION "Experiment 40a: PDITC iRIf slides" [115335-115703] -->
<h3 class="sectionedit305">2015.08.03</h3>
<div class="level3">
</div>
<h4>Experiment/Protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> 4 PDITC iRIf slides were prepared according to protocol</div>
</li>
<li class="level1"><div class="li"> plasma activation for 5 min at 80 L/h</div>
</li>
<li class="level1"><div class="li"> slides were incubated in APTES for 45 min</div>
</li>
<li class="level1"><div class="li"> slides were incubated in PDITC for 2 h, washed with Ethanol and Acetone and stored in the desiccator for 40 min</div>
</li>
<li class="level1"><div class="li"> 3µL of the Antibodies for HIV, HCV, Tetanus and Salmonella + controls (GFP, bBSA) were spotted</div>
</li>
</ul>
<p>
spotting pattern:
<a class="media" href="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" title="labjournal:surchem:surchem_11_slide_usage.png"><img alt="" class="media" src="https://static.igem.org/mediawiki/2015/4/4e/Freiburg_labjournal-surchem-surchem_11_slide_usage.png" width="300"/></a>
</p>
<div class="table sectionedit306"><table class="inline">
<tr class="row0">
<th class="col0"> # </th><th class="col1"> spot (AB) </th><th class="col2"> AB-Concentration [µg/mL] </th><th class="col3"> Antigen </th><th class="col4"> Elution no. </th><th class="col5"> Antigenconc. </th>
</tr>
<tr class="row1">
<td class="col0"> 1-2 </td><td class="col1"> gp41 DDX1306 </td><td class="col2"> 100 </td><td class="col3"> HIV(17) </td><td class="col4"> 1 </td><td class="col5"> 1.0 mg/mL?? </td>
</tr>
<tr class="row2">
<td class="col0"> 3-4 </td><td class="col1"> HCV-AB </td><td class="col2"> 100 </td><td class="col3"> HCV(10) </td><td class="col4"> 1 </td><td class="col5"> 0.74 mg/mL?? </td>
</tr>
<tr class="row3">
<td class="col0"> 5-6 </td><td class="col1"> HYB 278-01 </td><td class="col2"> 100 </td><td class="col3"> Tetanus(11) </td><td class="col4"> 1 </td><td class="col5"> 3.75 mg/mL?? </td>
</tr>
<tr class="row4">
<td class="col0"> 7-8 </td><td class="col1"> GFP </td><td class="col2"> 1.0 mg/mL </td><td class="col3"> a-GFP (goat;biotinylated) </td><td class="col4 leftalign"> </td><td class="col5"> 5 </td>
</tr>
<tr class="row5">
<td class="col0"> 9 </td><td class="col1"> bBSA </td><td class="col2"> 0.1 mg/mL </td><td class="col3">Strep-Cy5 </td><td class="col4"> </td><td class="col5"> (strep-Cy5) 10 </td>
</tr>
<tr class="row6">
<td class="col0"> 10-11 </td><td class="col1"> a-Salmonella-(pIG15_1301) </td><td class="col2"> 100 </td><td class="col3"> Salmonella(15) </td><td class="col4"> 2 </td><td class="col5"> 6.7 mg/mL </td>
</tr>
</table></div>
<!-- EDIT306 TABLE [116198-116644] --><ul>
<li class="level1"><div class="li"> slides were incubated at 4°C o/n</div>
</li>
</ul>
</div>
<!-- EDIT305 SECTION "2015.08.03" [115704-116684] -->
<h3 class="sectionedit307">2015.08.04</h3>
<div class="level3">
</div>
<h4>Experiment/protocol</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> spots were blocked 2x with 10 mg/mL BSA for 5 min</div>
</li>
<li class="level1"><div class="li"> slide was blocked in 10 mg/mL in BSA for 30 min</div>
</li>
<li class="level1"><div class="li"> <strong>slides were washed 1x with ddH<sub>2</sub>O</strong></div>
</li>
<li class="level1"><div class="li"> slides were measured in iRIf</div>
</li>
</ul>
</div>
<h4>Measurement</h4>
<div class="level4">
<p>
Only 1 slide was measured. Second slide syringe didnt suck solutions.
</p>
<div class="table sectionedit308"><table class="inline">
<tr class="row0">
<th class="col0"> Reagent </th><th class="col1"> Step </th><th class="col2"> Flow rate (ul/min) </th><th class="col3"> Priming time </th><th class="col4 leftalign"> Concentration  </th>
</tr>
<tr class="row1">
<td class="col0">Buffer</td><td class="col1">1</td><td class="col2">60</td><td class="col3">940</td><td class="col4">1x</td>
</tr>
<tr class="row2">
<td class="col0">BSA</td><td class="col1">2</td><td class="col2">30</td><td class="col3">900</td><td class="col4">10 mg/ml</td>
</tr>
<tr class="row3">
<td class="col0">Buffer</td><td class="col1">3</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row4">
<td class="col0">HIV (17)</td><td class="col1">4</td><td class="col2">20</td><td class="col3">900</td><td class="col4">0.25 mg/ml</td>
</tr>
<tr class="row5">
<td class="col0">Buffer</td><td class="col1">5</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row6">
<td class="col0">HCV (10)</td><td class="col1">6</td><td class="col2">20</td><td class="col3">900</td><td class="col4">0.25 mg/ml</td>
</tr>
<tr class="row7">
<td class="col0">Buffer</td><td class="col1">7</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row8">
<td class="col0">Tetanus (11)</td><td class="col1">8</td><td class="col2">20</td><td class="col3">900</td><td class="col4">0.25 mg/ml</td>
</tr>
<tr class="row9">
<td class="col0">Buffer</td><td class="col1">9</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row10">
<td class="col0">Anti-Mouse</td><td class="col1">10</td><td class="col2">20</td><td class="col3">900</td><td class="col4">5 ug/ml</td>
</tr>
<tr class="row11">
<td class="col0">Buffer</td><td class="col1">11</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
<tr class="row12">
<td class="col0">Anti-GFP</td><td class="col1">12</td><td class="col2">20</td><td class="col3">900</td><td class="col4">3 ug/ml</td>
</tr>
<tr class="row13">
<td class="col0">Buffer</td><td class="col1">13</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1x</td>
</tr>
</table></div>
<!-- EDIT308 TABLE [117017-117418] -->
</div>
<h4>Results</h4>
<div class="level4">
<ul>
<li class="level1"><div class="li"> Antigens didn't bind to the immobilized antibodies, anti-mouse only bound to a-Tetanus spot, and not to a-HIV and a-HCV spots, even though they are from mice, too.</div>
</li>
</ul>
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/b/b0/Freiburg_labjournal-roi_selection_20150804_29.png" title="labjournal:roi_selection_20150804_29.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/b/b0/Freiburg_labjournal-roi_selection_20150804_29.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/b/b0/Freiburg_labjournal-roi_selection_20150804_29.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 29: Selection of ROIs</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/7/74/Freiburg_labjournal-bindingcurve_20150804_29_2.png" title="labjournal:bindingcurve_20150804_29_2.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/74/Freiburg_labjournal-bindingcurve_20150804_29_2.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/7/74/Freiburg_labjournal-bindingcurve_20150804_29_2.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 29: Binding curve</div></div></div>
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