Difference between revisions of "Team:Freiburg/Labjournals/SurChem/September"

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<h1 class="sectionedit1">Labjournal Surchem September</h1>
 
<h1 class="sectionedit1">Labjournal Surchem September</h1>
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</div>
 
<!-- EDIT1 SECTION "Labjournal Surchem September" [1-43] -->
 
<!-- EDIT1 SECTION "Labjournal Surchem September" [1-43] -->
−
<h2 class="sectionedit2">Experiment 65b:   PDITC slides for iRIf</h2>
+
<h2 class="sectionedit2">Experiment 68a: Tetanus antigen on  PDITC slides IX</h2>
 
<div class="level2">
 
<div class="level2">
 
<ul>
 
<ul>
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<!-- EDIT2 SECTION "Experiment 65b:  PDITC slides  for iRIf" [44-218] -->
+
<!-- EDIT2 SECTION "Experiment 68a:  Tetanus antigen on  PDITC slides IX" [44-232] -->
−
<h3 class="sectionedit3">2015.09.04</h3>
+
<h3 class="sectionedit3">2015.09.16</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"> desiccator: 15 min</div>
 +
</li>
 +
</ul>
 +
</div>
 +
<!-- EDIT3 SECTION "2015.09.16" [233-430] -->
 +
<h3 class="sectionedit4">2015.09.16</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/2015/0/09/Freiburg_labjournal-surchem-spotting_pattern_8.png" width="300"/><br/>
 +
</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"> pos. control (GFP) </td><td class="col2"> 0.5 mg/ml </td>
 +
</tr>
 +
<tr class="row2">
 +
<td class="col0"> 2 </td><td class="col1"> BSA </td><td class="col2"> 5 mg/ml </td>
 +
</tr>
 +
<tr class="row3">
 +
<td class="col0"> 3 </td><td class="col1 leftalign"> Tetanus Antigen  </td><td class="col2"> ~0.5 mg/ml </td>
 +
</tr>
 +
<tr class="row4">
 +
<td class="col0"> 4 </td><td class="col1"> pos. control (GFP) </td><td class="col2"> 0.5 mg/ml </td>
 +
</tr>
 +
<tr class="row5">
 +
<td class="col0"> 5 </td><td class="col1"> BSA </td><td class="col2"> 5 mg/ml </td>
 +
</tr>
 +
<tr class="row6">
 +
<td class="col0"> 6 </td><td class="col1 leftalign"> Tetanus Antigen  </td><td class="col2"> ~0.5 mg/ml </td>
 +
</tr>
 +
<tr class="row7">
 +
<td class="col0"> 7 </td><td class="col1"> pos. control (GFP) </td><td class="col2"> 0.5 mg/ml </td>
 +
</tr>
 +
<tr class="row8">
 +
<td class="col0"> 8 </td><td class="col1 leftalign"> Tetanus Antigen  </td><td class="col2"> ~0.5 mg/ml </td>
 +
</tr>
 +
</table></div>
 +
<!-- EDIT5 TABLE [560-871] -->
 +
<p>
 +
  * spots were incubated o/n at 4 °C
 +
</p>
 +
</div>
 +
<!-- EDIT4 SECTION "2015.09.16" [431-910] -->
 +
<h3 class="sectionedit6">2015.09.17</h3>
 +
<div class="level3">
 +
</div>
 +
<h4>Experiment/Protocol</h4>
 +
<div class="level4">
 +
<p>
 +
Tet serum (-+): 024, 466 <br/>
 +
</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">300</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">Serum(-)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">900</td><td class="col4">1:10</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 GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">900</td><td class="col4">1.5 µg/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">Serum(-)</td><td class="col1">8</td><td class="col2">30</td><td class="col3">900</td><td class="col4">1:10</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 [989-1269] -->
 +
<p>
 +
Tet serum (+-): 208, 216 <br/>
 +
</p>
 +
<div class="table sectionedit8"><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">300</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">Serum(-)</td><td class="col1">4</td><td class="col2">30</td><td class="col3">900</td><td class="col4">1:10</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 GFP</td><td class="col1">6</td><td class="col2">30</td><td class="col3">900</td><td class="col4">1.5 µg/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">Serum(+)</td><td class="col1">8</td><td class="col2">30</td><td class="col3">900</td><td class="col4">1:10</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>
 +
<!-- EDIT8 TABLE [1301-1581] -->
 +
<p>
 +
GFP; Tet serum (-+): 287 <br/>
 +
</p>
 +
<div class="table sectionedit9"><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">300</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">900</td><td class="col4">1.5 µg/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(-)</td><td class="col1">6</td><td class="col2">30</td><td class="col3">900</td><td class="col4">1:10</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">Serum(+)</td><td class="col1">8</td><td class="col2">30</td><td class="col3">900</td><td class="col4">1:10</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>
 +
<!-- EDIT9 TABLE [1613-1893] -->
 +
</div>
 +
<h4>Results</h4>
 +
<div class="level4">
 +
<p>
 +
Serum(-),anti GFP,Serum(+):
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/45/Freiburg_labjournal-surchem-exp_68a_slide024.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_68a_slide024.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 024: Quotion picture, flushed with 1:10 diluted blood serum, flush protocol: serum(-),anti GFP, serum(+)</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/0/0b/Freiburg_labjournal-surchem-exp_68a_slide024_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_68a_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>
 +
<p>
 +
]
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/1/14/Freiburg_labjournal-surchem-exp_68a_slide466.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_68a_slide466.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 466: Quotion picture, flushed with 1:10 diluted blood serum, flush protocol: serum(-),anti GFP, serum(+)</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/49/Freiburg_labjournal-surchem-exp_68a_slide466_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_68a_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><hr/>
 +
<p>
 +
Serum(+),anti GFP,Serum(-):
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/d/d0/Freiburg_labjournal-surchem-exp_68a_slide208.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_68a_slide208.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 208: Quotion picture, flushed with 1:10 diluted blood serum, flush protocol: serum(+),anti GFP, serum(-)</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/e/e1/Freiburg_labjournal-surchem-exp_68a_slide208_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_68a_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><div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/f4/Freiburg_labjournal-surchem-exp_68a_slide216.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_68a_slide216.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 216: Quotion picture, flushed with 1:10 diluted blood serum, flush protocol: serum(+),anti GFP, serum(-)</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/d/d6/Freiburg_labjournal-surchem-exp_68a_slide216_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_68a_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><hr/>
 +
<p>
 +
anti GFP,Serum(-),Serum(+):
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/40/Freiburg_labjournal-surchem-exp_68a_slide287.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_68a_slide287.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 287: Quotion picture, flushed with 1:10 diluted blood serum, flush protocol: anti GFP, serum(-), serum(+)</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/f0/Freiburg_labjournal-surchem-exp_68a_slide287_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_68a_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>
 +
</div>
 +
<!-- EDIT6 SECTION "2015.09.17" [911-3321] -->
 +
<h2 class="sectionedit10">Experiment 67b:  Tetanus antigen on  PDITC slides VIII</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>
 +
<!-- EDIT10 SECTION "Experiment 67b:  Tetanus antigen on  PDITC slides VIII" [3322-3512] -->
 +
<h3 class="sectionedit11">2015.09.12</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"> desiccator: 15 min</div>
 +
</li>
 +
</ul>
 +
</div>
 +
<!-- EDIT11 SECTION "2015.09.12" [3513-3710] -->
 +
<h3 class="sectionedit12">2015.09.13</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/d/dd/Freiburg_labjournal-surchem-spotting_pattern6.png" width="300"/><br/>
 +
</p>
 +
<div class="table sectionedit13"><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"> BSA </td><td class="col2"> 5 mg/ml </td>
 +
</tr>
 +
<tr class="row3">
 +
<td class="col0"> 3 </td><td class="col1 leftalign"> Tetanus Antigen  </td><td class="col2"> ~0.5 mg/ml </td>
 +
</tr>
 +
<tr class="row4">
 +
<td class="col0"> 4 </td><td class="col1"> - </td><td class="col2"> - </td>
 +
</tr>
 +
<tr class="row5">
 +
<td class="col0"> 5 </td><td class="col1">-</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>
 +
</table></div>
 +
<!-- EDIT13 TABLE [3839-4022] -->
 +
<p>
 +
  * spots were incubated o/n at 4 °C
 +
</p>
 +
</div>
 +
<!-- EDIT12 SECTION "2015.09.13" [3711-4061] -->
 +
<h3 class="sectionedit14">2015.09.14</h3>
 +
<div class="level3">
 +
</div>
 +
<h4>Experiment/Protocol</h4>
 +
<div class="level4">
 +
<p>
 +
Tet serum (-): 208, 254 <br/>
 +
 
 +
Tet serum (+): 500, 024
 +
</p>
 +
<div class="table sectionedit15"><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">1.5 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">Serum SIG002 (-)/(+)</td><td class="col1">6</td><td class="col2">30</td><td class="col3">900</td><td class="col4">1:20</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 Human</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">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">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">60</td><td class="col3">300</td><td class="col4">1x</td>
 +
</tr>
 +
</table></div>
 +
<!-- EDIT15 TABLE [4163-4510] -->
 +
</div>
 +
<h4>Results</h4>
 +
<div class="level4">
 +
<p>
 +
Blood Serum(-):
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/3/3c/Freiburg_labjournal-surchem-exp_67b_slide208.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67b_slide208.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 208: Quotion picture, flushed with 1:20 diluted blood serum (-)</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/c/c7/Freiburg_labjournal-surchem-exp_67b_slide208_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67b_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>
 +
]
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/3/3b/Freiburg_labjournal-surchem-exp_67b_slide254.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67b_slide254.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 254: Quotion picture, flushed with 1:20 diluted blood serum (-)</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/e/e0/Freiburg_labjournal-surchem-exp_67b_slide254_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67b_slide254_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><hr/>
 +
<p>
 +
Blood serum(+):
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/a/a7/Freiburg_labjournal-surchem-exp_67b_slide024.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67b_slide024.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 024: Quotion picture, flushed with 1:20 diluted blood serum (+)</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/a/af/Freiburg_labjournal-surchem-exp_67b_slide024_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67b_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/2015/2/29/Freiburg_labjournal-surchem-exp_67b_slide500.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67b_slide500.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 500: Quotion picture, flushed with 1:20 diluted blood serum (+)</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/5/50/Freiburg_labjournal-surchem-exp_67b_slide500_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67b_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>
 +
</div>
 +
<!-- EDIT14 SECTION "2015.09.14" [4062-5455] -->
 +
<h2 class="sectionedit16">Experiment 67a:  Tetanus antigen on  PDITC slides VII</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>
 +
<!-- EDIT16 SECTION "Experiment 67a:  Tetanus antigen on  PDITC slides VII" [5456-5645] -->
 +
<h3 class="sectionedit17">2015.09.12</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"> desiccator: 15 min</div>
 +
</li>
 +
</ul>
 +
<p>
 +
Spotting pattern: <img alt="" class="media" src="https://static.igem.org/mediawiki/2015/d/dd/Freiburg_labjournal-surchem-spotting_pattern6.png" width="300"/><br/>
 +
</p>
 +
<div class="table sectionedit18"><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"> BSA </td><td class="col2"> 5 mg/ml </td>
 +
</tr>
 +
<tr class="row3">
 +
<td class="col0"> 3 </td><td class="col1 leftalign"> Tetanus Antigen  </td><td class="col2"> ~0.5 mg/ml </td>
 +
</tr>
 +
<tr class="row4">
 +
<td class="col0"> 4 </td><td class="col1"> - </td><td class="col2"> - </td>
 +
</tr>
 +
<tr class="row5">
 +
<td class="col0"> 5 </td><td class="col1">-</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>
 +
</table></div>
 +
<!-- EDIT18 TABLE [5924-6107] -->
 +
<p>
 +
  * spots were incubated o/n at 4 °C
 +
</p>
 +
</div>
 +
<!-- EDIT17 SECTION "2015.09.12" [5646-6146] -->
 +
<h3 class="sectionedit19">2015.09.13</h3>
 +
<div class="level3">
 +
</div>
 +
<h4>Experiment/Protocol</h4>
 +
<div class="level4">
 +
<p>
 +
Tet serum (-): 443, 466 <br/>
 +
 
 +
Tet serum (+): 423, 424
 +
</p>
 +
<div class="table sectionedit20"><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">1.5 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">Serum SIG002 (-)/(+)</td><td class="col1">6</td><td class="col2">30</td><td class="col3">900</td><td class="col4">1:10</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 Human</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">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">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">60</td><td class="col3">300</td><td class="col4">1x</td>
 +
</tr>
 +
</table></div>
 +
<!-- EDIT20 TABLE [6248-6595] -->
 +
</div>
 +
<h4>Results</h4>
 +
<div class="level4">
 +
<p>
 +
Blood Serum(-):
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/2e/Freiburg_labjournal-surchem-exp_67a_slide443.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67a_slide443.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 443: Quotion picture, flushed with 1:10 diluted blood serum (-)</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/4c/Freiburg_labjournal-surchem-exp_67a_slide443_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67a_slide443_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 443: binding curves</div></div></div>
 +
<p>
 +
]
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/74/Freiburg_labjournal-surchem-exp_67a_slide466.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67a_slide466.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 466: Quotion picture, flushed with 1:10 diluted blood serum (-)</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/8/81/Freiburg_labjournal-surchem-exp_67a_slide466_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67a_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><hr/>
 +
<p>
 +
Blood serum(+):
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/6b/Freiburg_labjournal-surchem-exp_67a_slide423.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67a_slide423.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 423: Quotion picture, flushed with 1:10 diluted blood serum (+)</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/c/c8/Freiburg_labjournal-surchem-exp_67a_slide423_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67a_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><div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/d/db/Freiburg_labjournal-surchem-exp_67a_slide424.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67a_slide424.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 424: Quotion picture, flushed with 1:10 diluted blood serum (+)</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/a/ab/Freiburg_labjournal-surchem-exp_67a_slide424_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_67a_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>
 +
</div>
 +
<!-- EDIT19 SECTION "2015.09.13" [6147-7540] -->
 +
<h2 class="sectionedit21">Experiment 66b:  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>
 +
<!-- EDIT21 SECTION "Experiment 66b:  PDITC slides  for iRIf" [7541-7715] -->
 +
<h3 class="sectionedit22">2015.09.06</h3>
 +
<div class="level3">
 +
</div>
 +
<h4>Experiment/Protocol</h4>
 +
<div class="level4">
 +
<ul>
 +
<li class="level1"><div class="li"> 6 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>
 +
<!-- EDIT22 SECTION "2015.09.06" [7716-7933] -->
 +
<h2 class="sectionedit23">Experiment 66a:  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>
 +
<!-- EDIT23 SECTION "Experiment 66a:  PDITC slides  for iRIf" [7934-8108] -->
 +
<h3 class="sectionedit24">2015.09.06</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"> desiccator: 15 min</div>
 +
</li>
 +
<li class="level1"><div class="li"> stored at 4 °C</div>
 +
</li>
 +
</ul>
 +
</div>
 +
<!-- EDIT24 SECTION "2015.09.06" [8109-8326] -->
 +
<h3 class="sectionedit25">2015.09.10</h3>
 +
<div class="level3">
 +
</div>
 +
<h4>Experiment/Protocol</h4>
 +
<div class="level4">
 +
<ul>
 +
<li class="level1"><div class="li"> slides were sandwiched with NTA and incubated at 4°C o/n</div>
 +
</li>
 +
</ul>
 +
</div>
 +
<!-- EDIT25 SECTION "2015.09.10" [8327-8438] -->
 +
<h3 class="sectionedit26">2015.09.10</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>
 +
<p>
 +
<p><div class="noteimportant">uploaded 20150906
 +
</div></p>
 +
</p>
 +
</div>
 +
<!-- EDIT26 SECTION "2015.09.10" [8439-8749] -->
 +
<h2 class="sectionedit27">Experiment 65c:  Tetanus antigen PDITC slides  for iRIf VI</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>
 +
<!-- EDIT27 SECTION "Experiment 65c:  Tetanus antigen PDITC slides  for iRIf VI" [8750-8942] -->
 +
<h3 class="sectionedit28">2015.08.31</h3>
 
<div class="level3">
 
<div class="level3">
 
</div>
 
</div>
Line 67: Line 587:
 
</ul>
 
</ul>
 
</div>
 
</div>
−
<!-- EDIT3 SECTION "2015.09.04" [219-436] -->
+
<!-- EDIT28 SECTION "2015.08.31" [8943-9160] -->
−
<h2 class="sectionedit4">Experiment 65b:  Tetanus antigen PDITC slides  for iRIf V</h2>
+
<h3 class="sectionedit29">2015.09.05</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/d/dd/Freiburg_labjournal-surchem-spotting_pattern6.png" width="300"/><br/>
 +
</p>
 +
<div class="table sectionedit30"><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"> BSA </td><td class="col2"> 5 mg/ml </td>
 +
</tr>
 +
<tr class="row3">
 +
<td class="col0"> 3 </td><td class="col1 leftalign"> Tetanus Antigen  </td><td class="col2"> ~0.5 mg/ml </td>
 +
</tr>
 +
<tr class="row4">
 +
<td class="col0"> 4 </td><td class="col1"> - </td><td class="col2"> - </td>
 +
</tr>
 +
<tr class="row5">
 +
<td class="col0"> 5 </td><td class="col1">-</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>
 +
</table></div>
 +
<!-- EDIT30 TABLE [9290-9473] -->
 +
</div>
 +
<!-- EDIT29 SECTION "2015.09.05" [9161-9475] -->
 +
<h3 class="sectionedit31">2015.09.06</h3>
 +
<div class="level3">
 +
</div>
 +
<h4>Experiment/Protocol</h4>
 +
<div class="level4">
 +
<p>
 +
Flush protocol:
 +
</p>
 +
<p>
 +
Tet serum (-): 443 <br/>
 +
 
 +
Tet serum (+): 466
 +
</p>
 +
<div class="table sectionedit32"><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">1.5 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">Serum SIG002 (-)/(+)</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1:4</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">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">300</td><td class="col4">1x</td>
 +
</tr>
 +
</table></div>
 +
<!-- EDIT32 TABLE [9585-9878] -->
 +
<p>
 +
Tet serum (-): 208<br/>
 +
 
 +
Tet serum (+): 303
 +
</p>
 +
<div class="table sectionedit33"><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">1.5 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">Serum SIG002 (-)/(+)</td><td class="col1">6</td><td class="col2">60</td><td class="col3">600</td><td class="col4">1:6</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">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">300</td><td class="col4">1x</td>
 +
</tr>
 +
</table></div>
 +
<!-- EDIT33 TABLE [9922-10215] -->
 +
<p>
 +
Note: the amount of flushed serum
 +
</p>
 +
</div>
 +
<h4>Results</h4>
 +
<div class="level4">
 +
<p>
 +
Blood Serum(-):
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/a/a1/Freiburg_labjournal-surchem-exp_65b_slide208.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65b_slide208.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 208: Quotion picture, flushed with 1:6 diluted blood serum (-)</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/77/Freiburg_labjournal-surchem-exp_65b_slide208_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65b_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>
 +
]
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/65/Freiburg_labjournal-surchem-exp_65b_slide443.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65b_slide443.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 443: Quotion picture, flushed with 1:4 diluted blood serum (-)</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/1/16/Freiburg_labjournal-surchem-exp_65b_slide443_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65b_slide443_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 443: binding curves</div></div></div><hr/>
 +
<p>
 +
Blood serum(+):
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/9/95/Freiburg_labjournal-surchem-exp_65b_slide303.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65b_slide303.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 303: Quotion picture, flushed with 1:6 diluted blood serum (+)</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/fc/Freiburg_labjournal-surchem-exp_65b_slide303_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65b_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 class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/0/06/Freiburg_labjournal-surchem-exp_65b_slide466.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65b_slide466.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 466: Quotion picture, flushed with 1:4 diluted blood serum (+)</div></div></div><div class="thumb2 trien" style="width:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/a/ac/Freiburg_labjournal-surchem-exp_65b_slide466_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65b_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>
 +
→ for the blood serum after immunization (+) the iRIf signal for tetanus binding is higher than for the blood serum before immunization (-)
 +
</p>
 +
<p>
 +
→ there are differences to the results of experiment 59a, the overall tetanus signal is much weaker as well as the binding of the blood serum to bBSA
 +
</p>
 +
</div>
 +
<!-- EDIT31 SECTION "2015.09.06" [9476-11483] -->
 +
<h2 class="sectionedit34">Experiment 65b:  Tetanus antigen PDITC slides  for iRIf V</h2>
 
<div class="level2">
 
<div class="level2">
 
<ul>
 
<ul>
Line 75: Line 739:
 
</ul>
 
</ul>
 
</div>
 
</div>
−
<!-- EDIT4 SECTION "Experiment 65b:  Tetanus antigen PDITC slides  for iRIf V" [437-627] -->
+
<!-- EDIT34 SECTION "Experiment 65b:  Tetanus antigen PDITC slides  for iRIf V" [11484-11674] -->
−
<h3 class="sectionedit5">2015.09.04</h3>
+
<h3 class="sectionedit35">2015.08.31</h3>
 
<div class="level3">
 
<div class="level3">
 
</div>
 
</div>
Line 93: Line 757:
 
</li>
 
</li>
 
</ul>
 
</ul>
 +
</div>
 +
<!-- EDIT35 SECTION "2015.08.31" [11675-11872] -->
 +
<h3 class="sectionedit36">2015.09.04</h3>
 +
<div class="level3">
 +
</div>
 +
<h4>Experiment/Protocol</h4>
 +
<div class="level4">
 
<p>
 
<p>
 
Spotting pattern: <img alt="" class="media" src="https://static.igem.org/mediawiki/2015/d/dd/Freiburg_labjournal-surchem-spotting_pattern6.png" width="300"/><br/>
 
Spotting pattern: <img alt="" class="media" src="https://static.igem.org/mediawiki/2015/d/dd/Freiburg_labjournal-surchem-spotting_pattern6.png" width="300"/><br/>
 
</p>
 
</p>
−
<div class="table sectionedit6"><table class="inline">
+
<div class="table sectionedit37"><table class="inline">
 
<tr class="row0">
 
<tr class="row0">
 
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
 
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
Line 119: Line 790:
 
</tr>
 
</tr>
 
</table></div>
 
</table></div>
−
<!-- EDIT6 TABLE [905-1088] -->
+
<!-- EDIT37 TABLE [12001-12184] -->
 
</div>
 
</div>
−
<!-- EDIT5 SECTION "2015.09.04" [628-1089] -->
+
<!-- EDIT36 SECTION "2015.09.04" [11873-12185] -->
−
<h3 class="sectionedit7">2015.09.05</h3>
+
<h3 class="sectionedit38">2015.09.05</h3>
 
<div class="level3">
 
<div class="level3">
 
</div>
 
</div>
Line 131: Line 802:
 
</p>
 
</p>
 
<p>
 
<p>
−
Tet serum (-): <br/>
+
SIG 003 (+): 012(no serum was flushed over the slide), 315, 426<br/>
  
−
Tet serum (+):  
+
SIG 002 (+): 215
 
</p>
 
</p>
−
<div class="table sectionedit8"><table class="inline">
+
<div class="table sectionedit39"><table class="inline">
 
<tr class="row0">
 
<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>
 
<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>
Line 155: Line 826:
 
</tr>
 
</tr>
 
<tr class="row6">
 
<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">1:10/1:30</td>
+
<td class="col0">Blood serum</td><td class="col1">6</td><td class="col2">30</td><td class="col3">600</td><td class="col4">1:30</td>
 
</tr>
 
</tr>
 
<tr class="row7">
 
<tr class="row7">
Line 167: Line 838:
 
</tr>
 
</tr>
 
</table></div>
 
</table></div>
−
<!-- EDIT8 TABLE [1191-1490] -->
+
<!-- EDIT39 TABLE [12336-12621] -->
 +
</div>
 +
<h4>Results</h4>
 +
<div class="level4">
 +
<p>
 +
SIG 003:
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/28/Freiburg_labjournal-surchem-exp_65b_slide012.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65b_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:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/74/Freiburg_labjournal-surchem-exp_65b_slide012_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65b_slide012_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><div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/f1/Freiburg_labjournal-surchem-exp_65b_slide315.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65b_slide315.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 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/4/46/Freiburg_labjournal-surchem-exp_65b_slide426.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65b_slide426.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 426: 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/2015/3/31/Freiburg_labjournal-surchem-exp_65b_slide426_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65b_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>
 +
<p>
 +
→ slide 012 was not flushed with any blood serum
 +
→ for blood serum SIG 003 only a slight antigen/antibody binding is detectable (slide 426)
 +
</p>
 +
<hr/>
 +
<p>
 +
SIG 002:
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/e/eb/Freiburg_labjournal-surchem-exp_65b_slide215.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65b_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:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/9/93/Freiburg_labjournal-surchem-exp_65b_slide215_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65b_slide215_binding.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>
 +
<p>
 +
→ slight antigen/antibody binding is detectable
 +
</p>
 +
<p>
 +
→ no anti GFP/ GFP binding detected
 +
</p>
 +
<p>
 +
→ only weak binding of the blood serum to bBSA → difference to results of experiment 59a
 +
</p>
 
</div>
 
</div>
−
<!-- EDIT7 SECTION "2015.09.05" [1090-1492] -->
+
<!-- EDIT38 SECTION "2015.09.05" [12186-13605] -->
−
<h2 class="sectionedit9">Experiment 65a: cell free GFP on NiNTA and PDITC slides  for iRIf</h2>
+
<h2 class="sectionedit40">Experiment 65a: cell free GFP on NiNTA and PDITC slides  for iRIf</h2>
 
<div class="level2">
 
<div class="level2">
 
<ul>
 
<ul>
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</ul>
 
</ul>
 
</div>
 
</div>
−
<!-- EDIT9 SECTION "Experiment 65a: cell free GFP on NiNTA and PDITC slides  for iRIf" [1493-1692] -->
+
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−
<h3 class="sectionedit10">2015.09.04</h3>
+
<h3 class="sectionedit41">2015.08.31</h3>
 
<div class="level3">
 
<div class="level3">
 
</div>
 
</div>
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</li>
 
</li>
 
</ul>
 
</ul>
 +
</div>
 +
<!-- EDIT41 SECTION "2015.08.31" [13806-14081] -->
 +
<h3 class="sectionedit42">2015.09.04</h3>
 +
<div class="level3">
 +
</div>
 +
<h4>Experiment/Protocol</h4>
 +
<div class="level4">
 
<p>
 
<p>
 
Spotting pattern:<img alt="" class="media" src="https://static.igem.org/mediawiki/2015/0/09/Freiburg_labjournal-surchem-spotting_pattern_8.png" width="300"/><br/>
 
Spotting pattern:<img alt="" class="media" src="https://static.igem.org/mediawiki/2015/0/09/Freiburg_labjournal-surchem-spotting_pattern_8.png" width="300"/><br/>
 
</p>
 
</p>
−
<div class="table sectionedit11"><table class="inline">
+
<div class="table sectionedit43"><table class="inline">
 
<tr class="row0">
 
<tr class="row0">
 
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
 
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
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</tr>
 
</tr>
 
</table></div>
 
</table></div>
−
<!-- EDIT11 TABLE [2048-2405] -->
+
<!-- EDIT43 TABLE [14210-14567] -->
 
</div>
 
</div>
−
<!-- EDIT10 SECTION "2015.09.04" [1693-2406] -->
+
<!-- EDIT42 SECTION "2015.09.04" [14082-14568] -->
−
<h3 class="sectionedit12">2015.09.05</h3>
+
<h3 class="sectionedit44">2015.09.05</h3>
 
<div class="level3">
 
<div class="level3">
 
</div>
 
</div>
Line 240: Line 943:
 
Flush protocol:
 
Flush protocol:
 
</p>
 
</p>
−
<div class="table sectionedit13"><table class="inline">
+
<div class="table sectionedit45"><table class="inline">
 
<tr class="row0">
 
<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>
 
<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>
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</tr>
 
</tr>
 
</table></div>
 
</table></div>
−
<!-- EDIT13 TABLE [2473-2709] -->
+
<!-- EDIT45 TABLE [14635-14871] -->
 +
</div>
 +
<h4>Results</h4>
 +
<div class="level4">
 +
<p>
 +
PDITC:
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/f7/Freiburg_labjournal-surchem-exp_65a_slide216.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65a_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:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/f/ff/Freiburg_labjournal-surchem-exp_65a_slide216_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65a_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/d/d0/Freiburg_labjournal-surchem-exp_65a_slide254.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65a_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:510px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/72/Freiburg_labjournal-surchem-exp_65a_slide254_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65a_slide254_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><hr/>
 +
<p>
 +
Ni-NTA:
 +
</p>
 +
<div class="thumb2 tleft" style="width:310px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/3/35/Freiburg_labjournal-surchem-exp_65a_slide102.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65a_slide102.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 102: 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/2015/9/95/Freiburg_labjournal-surchem-exp_65a_slide102_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65a_slide102_binding.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 102: 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/f/fc/Freiburg_labjournal-surchem-exp_65a_slide502.png" width="300"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65a_slide502.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 502: 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/e/e7/Freiburg_labjournal-surchem-exp_65a_slide504_binding.png" width="500"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Asurchem%3Aseptember&amp;media=labjournal:surchem:exp_65a_slide504_binding.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>
 +
<p>
 +
→ the his tagged, cell free GFP was specificly bound by the Ni NTA surface, whereas there was no detectable amount of cell free GFP bound on the PDITC surfaces
 +
</p>
 +
<p>
 +
→ the PDITC surface binds all proteins in the spotted cell free expression mix eaqually good, whereas the Ni-NTA surface specificly binds only the his tagged, cell free expressed GFP
 +
</p>
 
</div>
 
</div>
−
<!-- EDIT12 SECTION "2015.09.05" [2407-2710] -->
+
<!-- EDIT44 SECTION "2015.09.05" [14569-15968] -->
−
<h2 class="sectionedit14">Experiment 64c:  Tetanus antigen PDITC slides  for iRIf IV</h2>
+
<h2 class="sectionedit46">Experiment 64c:  Tetanus antigen PDITC slides  for iRIf IV</h2>
 
<div class="level2">
 
<div class="level2">
 
<ul>
 
<ul>
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</ul>
 
</ul>
 
</div>
 
</div>
−
<!-- EDIT14 SECTION "Experiment 64c:  Tetanus antigen PDITC slides  for iRIf IV" [2711-2902] -->
+
<!-- EDIT46 SECTION "Experiment 64c:  Tetanus antigen PDITC slides  for iRIf IV" [15969-16160] -->
−
<h3 class="sectionedit15">2015.08.31</h3>
+
<h3 class="sectionedit47">2015.08.31</h3>
 
<div class="level3">
 
<div class="level3">
 
</div>
 
</div>
Line 297: Line 1,017:
 
</ul>
 
</ul>
 
</div>
 
</div>
−
<!-- EDIT15 SECTION "2015.08.31" [2903-3120] -->
+
<!-- EDIT47 SECTION "2015.08.31" [16161-16378] -->
−
<h3 class="sectionedit16">2015.09.03</h3>
+
<h3 class="sectionedit48">2015.09.03</h3>
 
<div class="level3">
 
<div class="level3">
 
</div>
 
</div>
Line 306: Line 1,026:
 
Spotting pattern: <img alt="" class="media" src="https://static.igem.org/mediawiki/2015/d/dd/Freiburg_labjournal-surchem-spotting_pattern6.png" width="300"/><br/>
 
Spotting pattern: <img alt="" class="media" src="https://static.igem.org/mediawiki/2015/d/dd/Freiburg_labjournal-surchem-spotting_pattern6.png" width="300"/><br/>
 
</p>
 
</p>
−
<div class="table sectionedit17"><table class="inline">
+
<div class="table sectionedit49"><table class="inline">
 
<tr class="row0">
 
<tr class="row0">
 
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
 
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
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</tr>
 
</tr>
 
</table></div>
 
</table></div>
−
<!-- EDIT17 TABLE [3249-3447] -->
+
<!-- EDIT49 TABLE [16507-16705] -->
 
</div>
 
</div>
−
<!-- EDIT16 SECTION "2015.09.03" [3121-3449] -->
+
<!-- EDIT48 SECTION "2015.09.03" [16379-16707] -->
−
<h3 class="sectionedit18">2015.09.04</h3>
+
<h3 class="sectionedit50">2015.09.04</h3>
 
<div class="level3">
 
<div class="level3">
 
</div>
 
</div>
Line 345: Line 1,065:
 
Tet serum (+): 024 (1:10 diluted serum), 504 (1:30 diluted serum)
 
Tet serum (+): 024 (1:10 diluted serum), 504 (1:30 diluted serum)
 
</p>
 
</p>
−
<div class="table sectionedit19"><table class="inline">
+
<div class="table sectionedit51"><table class="inline">
 
<tr class="row0">
 
<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>
 
<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>
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</tr>
 
</tr>
 
</table></div>
 
</table></div>
−
<!-- EDIT19 TABLE [3625-3924] -->
+
<!-- EDIT51 TABLE [16883-17182] -->
 
</div>
 
</div>
 
<h4>Results</h4>
 
<h4>Results</h4>
Line 397: Line 1,117:
 
</p>
 
</p>
 
</div>
 
</div>
−
<!-- EDIT18 SECTION "2015.09.04" [3450-4640] -->
+
<!-- EDIT50 SECTION "2015.09.04" [16708-17898] -->
−
<h2 class="sectionedit20">Experiment 64b: Tetanus antigen PDITC slides  for iRIf III</h2>
+
<h2 class="sectionedit52">Experiment 64b: Tetanus antigen PDITC slides  for iRIf III</h2>
 
<div class="level2">
 
<div class="level2">
 
<ul>
 
<ul>
Line 405: Line 1,125:
 
</ul>
 
</ul>
 
</div>
 
</div>
−
<!-- EDIT20 SECTION "Experiment 64b: Tetanus antigen PDITC slides  for iRIf III" [4641-4832] -->
+
<!-- EDIT52 SECTION "Experiment 64b: Tetanus antigen PDITC slides  for iRIf III" [17899-18090] -->
−
<h3 class="sectionedit21">2015.08.31</h3>
+
<h3 class="sectionedit53">2015.08.31</h3>
 
<div class="level3">
 
<div class="level3">
 
</div>
 
</div>
Line 426: Line 1,146:
 
</ul>
 
</ul>
 
</div>
 
</div>
−
<!-- EDIT21 SECTION "2015.08.31" [4833-5051] -->
+
<!-- EDIT53 SECTION "2015.08.31" [18091-18309] -->
−
<h3 class="sectionedit22">2015.09.02</h3>
+
<h3 class="sectionedit54">2015.09.02</h3>
 
<div class="level3">
 
<div class="level3">
 
</div>
 
</div>
Line 435: Line 1,155:
 
Spotting pattern: <img alt="" class="media" src="https://static.igem.org/mediawiki/2015/d/dd/Freiburg_labjournal-surchem-spotting_pattern6.png" width="300"/><br/>
 
Spotting pattern: <img alt="" class="media" src="https://static.igem.org/mediawiki/2015/d/dd/Freiburg_labjournal-surchem-spotting_pattern6.png" width="300"/><br/>
 
</p>
 
</p>
−
<div class="table sectionedit23"><table class="inline">
+
<div class="table sectionedit55"><table class="inline">
 
<tr class="row0">
 
<tr class="row0">
 
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
 
<th class="col0"> # </th><th class="col1 leftalign"> spot  </th><th class="col2"> Concentration </th>
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</tr>
 
</tr>
 
</table></div>
 
</table></div>
−
<!-- EDIT23 TABLE [5180-5394] -->
+
<!-- EDIT55 TABLE [18438-18652] -->
 
<p>
 
<p>
 
note: bBSA on spot 5 was incubated for only a minute and then replaced with BSA   
 
note: bBSA on spot 5 was incubated for only a minute and then replaced with BSA   
 
</p>
 
</p>
 
</div>
 
</div>
−
<!-- EDIT22 SECTION "2015.09.02" [5052-5478] -->
+
<!-- EDIT54 SECTION "2015.09.02" [18310-18736] -->
−
<h3 class="sectionedit24">2015.09.03</h3>
+
<h3 class="sectionedit56">2015.09.03</h3>
 
<div class="level3">
 
<div class="level3">
 
</div>
 
</div>
Line 477: Line 1,197:
 
Tet serum (+): 443 (1:10 diluted serum), 423 (1:30 diluted serum)
 
Tet serum (+): 443 (1:10 diluted serum), 423 (1:30 diluted serum)
 
</p>
 
</p>
−
<div class="table sectionedit25"><table class="inline">
+
<div class="table sectionedit57"><table class="inline">
 
<tr class="row0">
 
<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>
 
<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>
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</tr>
 
</tr>
 
</table></div>
 
</table></div>
−
<!-- EDIT25 TABLE [5680-5974] -->
+
<!-- EDIT57 TABLE [18938-19232] -->
 
</div>
 
</div>
 
<h4>Results</h4>
 
<h4>Results</h4>
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</p>
 
</p>
 
</div>
 
</div>
−
<!-- EDIT24 SECTION "2015.09.03" [5479-7388] -->
+
<!-- EDIT56 SECTION "2015.09.03" [18737-20646] -->
−
<h2 class="sectionedit26">Experiment 64a: Salmonella antigen on PDITC slides  for iRIf II</h2>
+
<h2 class="sectionedit58">Experiment 64a: Salmonella antigen on PDITC slides  for iRIf II</h2>
 
<div class="level2">
 
<div class="level2">
 
<ul>
 
<ul>
Line 543: Line 1,263:
 
</ul>
 
</ul>
 
</div>
 
</div>
−
<!-- EDIT26 SECTION "Experiment 64a: Salmonella antigen on PDITC slides  for iRIf II" [7389-7586] -->
+
<!-- EDIT58 SECTION "Experiment 64a: Salmonella antigen on PDITC slides  for iRIf II" [20647-20844] -->
−
<h3 class="sectionedit27">2015.08.31</h3>
+
<h3 class="sectionedit59">2015.08.31</h3>
 
<div class="level3">
 
<div class="level3">
 
</div>
 
</div>
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</ul>
 
</ul>
 
</div>
 
</div>
−
<!-- EDIT27 SECTION "2015.08.31" [7587-7804] -->
+
<!-- EDIT59 SECTION "2015.08.31" [20845-21062] -->
−
<h3 class="sectionedit28">2015.09.01</h3>
+
<h3 class="sectionedit60">2015.09.01</h3>
 
<div class="level3">
 
<div class="level3">
 
</div>
 
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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/>
 
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Slides (3): 208,215,216
 
Slides (3): 208,215,216
 
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Flush protocol:
 
Flush protocol:
 
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<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>
 
<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>
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last slide (208) changed protocol: only 1 ug/ml anti-GFP used; anti-Salmonella desalt was mixed 1:1 with anti-Salmonella lysate
 
last slide (208) changed protocol: only 1 ug/ml anti-GFP used; anti-Salmonella desalt was mixed 1:1 with anti-Salmonella lysate
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Note: Slide 208 had the best signal.
 
Note: Slide 208 had the best signal.
 
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Revision as of 16:37, 18 September 2015

""

Labjournal Surchem September

Experiment 68a: Tetanus antigen on PDITC slides IX

2015.09.16

Experiment/Protocol

  • 4 PDITC slides were prepared
  • Plasma activation: 40 L/h, 5 min
  • APTES incubation: 30 min
  • PDITC incubation: 2 h
  • desiccator: 15 min

2015.09.16

Experiment/Protocol

Spotting pattern:

# spot Concentration
1 pos. control (GFP) 0.5 mg/ml
2 BSA 5 mg/ml
3 Tetanus Antigen ~0.5 mg/ml
4 pos. control (GFP) 0.5 mg/ml
5 BSA 5 mg/ml
6 Tetanus Antigen ~0.5 mg/ml
7 pos. control (GFP) 0.5 mg/ml
8 Tetanus Antigen ~0.5 mg/ml

* spots were incubated o/n at 4 °C

2015.09.17

Experiment/Protocol

Tet serum (-+): 024, 466

Reagent Step Flow rate (ul/min) Priming time Concentration
Buffer1603001x
BSA26060010 mg/ml
Buffer3606001x
Serum(-)4309001:10
Buffer5603001x
anti GFP6309001.5 µg/ml
Buffer7603001x
Serum(-)8309001:10
Buffer9603001x

Tet serum (+-): 208, 216

Reagent Step Flow rate (ul/min) Priming time Concentration
Buffer1603001x
BSA26060010 mg/ml
Buffer3606001x
Serum(-)4309001:10
Buffer5603001x
anti GFP6309001.5 µg/ml
Buffer7603001x
Serum(+)8309001:10
Buffer9603001x

GFP; Tet serum (-+): 287

Reagent Step Flow rate (ul/min) Priming time Concentration
Buffer1603001x
BSA26060010 mg/ml
Buffer3606001x
anti GFP4309001.5 µg/ml
Buffer5603001x
Serum(-)6309001:10
Buffer7603001x
Serum(+)8309001:10
Buffer9603001x

Results

Serum(-),anti GFP,Serum(+):

Slide 024: Quotion picture, flushed with 1:10 diluted blood serum, flush protocol: serum(-),anti GFP, serum(+)
Slide 024: binding curves

]

Slide 466: Quotion picture, flushed with 1:10 diluted blood serum, flush protocol: serum(-),anti GFP, serum(+)
Slide 466: binding curves

Serum(+),anti GFP,Serum(-):

Slide 208: Quotion picture, flushed with 1:10 diluted blood serum, flush protocol: serum(+),anti GFP, serum(-)
Slide 208: binding curves
Slide 216: Quotion picture, flushed with 1:10 diluted blood serum, flush protocol: serum(+),anti GFP, serum(-)
Slide 216: binding curves

anti GFP,Serum(-),Serum(+):

Slide 287: Quotion picture, flushed with 1:10 diluted blood serum, flush protocol: anti GFP, serum(-), serum(+)
Slide 287: binding curves

Experiment 67b: Tetanus antigen on PDITC slides VIII

2015.09.12

Experiment/Protocol

  • 4 PDITC slides were prepared
  • Plasma activation: 40 L/h, 5 min
  • APTES incubation: 30 min
  • PDITC incubation: 2 h
  • desiccator: 15 min

2015.09.13

Experiment/Protocol

Spotting pattern:

# spot Concentration
1 pos. control (GFP) 0.5 mg/ml
2 BSA 5 mg/ml
3 Tetanus Antigen ~0.5 mg/ml
4 - -
5 - -
6 bBSA 200 µg/ml

* spots were incubated o/n at 4 °C

2015.09.14

Experiment/Protocol

Tet serum (-): 208, 254
Tet serum (+): 500, 024

Reagent Step Flow rate (ul/min) Priming time Concentration
Buffer1606001x
BSA26060010 mg/ml
Buffer3606001x
anti-GFP4306001.5 ug/ml
Buffer5603001x
Serum SIG002 (-)/(+)6309001:20
Buffer7603001x
Anti Human8306005 ug/ml
Buffer9603001x
StrepCy510306005 ug/ml
Buffer11603001x

Results

Blood Serum(-):

Slide 208: Quotion picture, flushed with 1:20 diluted blood serum (-)
Slide 208: binding curves

]

Slide 254: Quotion picture, flushed with 1:20 diluted blood serum (-)
Slide 254: binding curves

Blood serum(+):

Slide 024: Quotion picture, flushed with 1:20 diluted blood serum (+)
Slide 024: binding curves
Slide 500: Quotion picture, flushed with 1:20 diluted blood serum (+)
Slide 500: binding curves

Experiment 67a: Tetanus antigen on PDITC slides VII

2015.09.12

Experiment/Protocol

  • 4 PDITC slides were prepared
  • Plasma activation: 40 L/h, 5 min
  • APTES incubation: 30 min
  • PDITC incubation: 2 h
  • desiccator: 15 min

Spotting pattern:

# spot Concentration
1 pos. control (GFP) 0.5 mg/ml
2 BSA 5 mg/ml
3 Tetanus Antigen ~0.5 mg/ml
4 - -
5 - -
6 bBSA 200 µg/ml

* spots were incubated o/n at 4 °C

2015.09.13

Experiment/Protocol

Tet serum (-): 443, 466
Tet serum (+): 423, 424

Reagent Step Flow rate (ul/min) Priming time Concentration
Buffer1606001x
BSA26060010 mg/ml
Buffer3606001x
anti-GFP4306001.5 ug/ml
Buffer5603001x
Serum SIG002 (-)/(+)6309001:10
Buffer7603001x
Anti Human8306005 ug/ml
Buffer9603001x
StrepCy510306005 ug/ml
Buffer11603001x

Results

Blood Serum(-):

Slide 443: Quotion picture, flushed with 1:10 diluted blood serum (-)
Slide 443: binding curves

]

Slide 466: Quotion picture, flushed with 1:10 diluted blood serum (-)
Slide 466: binding curves

Blood serum(+):

Slide 423: Quotion picture, flushed with 1:10 diluted blood serum (+)
Slide 423: binding curves
Slide 424: Quotion picture, flushed with 1:10 diluted blood serum (+)
Slide 424: binding curves

Experiment 66b: PDITC slides for iRIf

2015.09.06

Experiment/Protocol

  • 6 PDITC slides were prepared
  • Plasma activation: 40 L/h, 5 min
  • APTES incubation: 30 min
  • PDITC incubation: 2 h
  • desiccator: 15 min
  • stored at 4 °C

Experiment 66a: PDITC slides for iRIf

2015.09.06

Experiment/Protocol

  • 4 PDITC slides were prepared
  • Plasma activation: 40 L/h, 5 min
  • APTES incubation: 30 min
  • PDITC incubation: 2 h
  • desiccator: 15 min
  • stored at 4 °C

2015.09.10

Experiment/Protocol

  • slides were sandwiched with NTA and incubated at 4°C o/n

2015.09.10

Experiment/Protocol

  • slides were blocked with APTES blocking solution for 1 h
  • slides were washed 2x with PBS
  • slides were incubated with NiSO4-solution for 1 h and 20 min
  • slides were washed with PBS/2x diluted PBS

uploaded 20150906

Experiment 65c: Tetanus antigen PDITC slides for iRIf VI

2015.08.31

Experiment/Protocol

  • 4 PDITC slides were prepared
  • Plasma activation: 40 L/h, 5 min
  • APTES incubation: 30 min
  • PDITC incubation: 2 h
  • desiccator: 30 min
  • stored at 4 °C

2015.09.05

Experiment/Protocol

Spotting pattern:

# spot Concentration
1 pos. control (GFP) 0.5 mg/ml
2 BSA 5 mg/ml
3 Tetanus Antigen ~0.5 mg/ml
4 - -
5 - -
6 bBSA 200 µg/ml

2015.09.06

Experiment/Protocol

Flush protocol:

Tet serum (-): 443
Tet serum (+): 466

Reagent Step Flow rate (ul/min) Priming time Concentration
Buffer1606001x
BSA26060010 mg/ml
Buffer3606001x
anti-GFP4306001.5 ug/ml
Buffer5603001x
Serum SIG002 (-)/(+)6306001:4
Buffer7603001x
StrepCy58306005 ug/ml
Buffer9603001x

Tet serum (-): 208
Tet serum (+): 303

Reagent Step Flow rate (ul/min) Priming time Concentration
Buffer1606001x
BSA26060010 mg/ml
Buffer3606001x
anti-GFP4306001.5 ug/ml
Buffer5603001x
Serum SIG002 (-)/(+)6606001:6
Buffer7603001x
StrepCy58306005 ug/ml
Buffer9603001x

Note: the amount of flushed serum

Results

Blood Serum(-):

Slide 208: Quotion picture, flushed with 1:6 diluted blood serum (-)
Slide 208: binding curves

]

Slide 443: Quotion picture, flushed with 1:4 diluted blood serum (-)
Slide 443: binding curves

Blood serum(+):

Slide 303: Quotion picture, flushed with 1:6 diluted blood serum (+)
Slide 303: binding curves
Slide 466: Quotion picture, flushed with 1:4 diluted blood serum (+)
Slide 466: binding curves

→ for the blood serum after immunization (+) the iRIf signal for tetanus binding is higher than for the blood serum before immunization (-)

→ there are differences to the results of experiment 59a, the overall tetanus signal is much weaker as well as the binding of the blood serum to bBSA

Experiment 65b: Tetanus antigen PDITC slides for iRIf V

2015.08.31

Experiment/Protocol

  • 4 PDITC slides were prepared
  • Plasma activation: 40 L/h, 5 min
  • APTES incubation: 30 min
  • PDITC incubation: 2 h
  • desiccator: 30 min

2015.09.04

Experiment/Protocol

Spotting pattern:

# spot Concentration
1 pos. control (GFP) 0.5 mg/ml
2 BSA 5 mg/ml
3 Tetanus Antigen ~0.5 mg/ml
4 - -
5 - -
6 bBSA 200 µg/ml

2015.09.05

Experiment/Protocol

Flush protocol:

SIG 003 (+): 012(no serum was flushed over the slide), 315, 426
SIG 002 (+): 215

Reagent Step Flow rate (ul/min) Priming time Concentration
Buffer1606001x
BSA26060010 mg/ml
Buffer3606001x
anti-GFP4306001.5 ug/ml
Buffer5603001x
Blood serum6306001:30
Buffer7603001x
StrepCy58306005 ug/ml
Buffer9603001x

Results

SIG 003:

Slide 012: Quotion picture
Slide 012: binding curves
Slide 315: Quotion picture
Slide 426: Quotion picture
Slide 426: binding curves

→ slide 012 was not flushed with any blood serum → for blood serum SIG 003 only a slight antigen/antibody binding is detectable (slide 426)

SIG 002:

Slide 215: Quotion picture
Slide 215: binding curves

→ slight antigen/antibody binding is detectable

→ no anti GFP/ GFP binding detected

→ only weak binding of the blood serum to bBSA → difference to results of experiment 59a

Experiment 65a: cell free GFP on NiNTA and PDITC slides for iRIf

2015.08.31

Experiment/Protocol

  • 2 PDITC slides were prepared (216,254)
  • Plasma activation: 40 L/h, 5 min
  • APTES incubation: 30 min
  • PDITC incubation: 2 h
  • desiccator: 30 min
  • 2 Ni NTA slides from experiment 63b were also spotted (102,502)

2015.09.04

Experiment/Protocol

Spotting pattern:

# spot Concentration
1 pos. control (GFP) 0.5 mg/ml
2 cell free expression mix without DNA (KK)
3 cell free expressed HA-GFP-His6-His6 (KK)
4 pos. control (GFP) 0.5 mg/ml
5 cell free expression mix without DNA (KK)
6 cell free expressed His10-GFP-Spy (KK)
7 BSA 5 mg/ml
8 bBSA 200 µg/ml

2015.09.05

Experiment/Protocol

Flush protocol:

Reagent Step Flow rate (ul/min) Priming time Concentration
Buffer1606001x
BSA26060010 mg/ml
Buffer3606001x
anti-GFP4306001.5 ug/ml
Buffer5603001x
StrepCy58306005 ug/ml
Buffer9603001x

Results

PDITC:

Slide 216: Quotion picture
Slide 216: binding curves
Slide 254: Quotion picture
Slide 254: binding curves

Ni-NTA:

Slide 102: Quotion picture
Slide 102: binding curves
Slide 502: Quotion picture
Slide 502: binding curves

→ the his tagged, cell free GFP was specificly bound by the Ni NTA surface, whereas there was no detectable amount of cell free GFP bound on the PDITC surfaces

→ the PDITC surface binds all proteins in the spotted cell free expression mix eaqually good, whereas the Ni-NTA surface specificly binds only the his tagged, cell free expressed GFP

Experiment 64c: Tetanus antigen PDITC slides for iRIf IV

2015.08.31

Experiment/Protocol

  • 4 PDITC slides were prepared
  • Plasma activation: 40 L/h, 5 min
  • APTES incubation: 30 min
  • PDITC incubation: 2 h
  • desiccator: 15 min
  • stored at 4 °C

2015.09.03

Experiment/Protocol

Spotting pattern:

# spot Concentration
1 pos. control (GFP) 0.5 mg/ml
2 BSA 5 mg/ml
3 Tetanus Antigen ~0.5 mg/ml
4 - -
5 mCherry 0.5 mg/ml
6 bBSA 200 µg/ml

2015.09.04

Experiment/Protocol

Flush protocol:

Tet serum (-): 500(1:10 diluted serum)
Tet serum (+): 024 (1:10 diluted serum), 504 (1:30 diluted serum)

Reagent Step Flow rate (ul/min) Priming time Concentration
Buffer1606001x
BSA26060010 mg/ml
Buffer3606001x
anti-GFP4306001.5 ug/ml
Buffer5603001x
Serum SIG002 (-)/(+)6306001:10/1:30
Buffer7603001x
StrepCy58306005 ug/ml
Buffer9603001x

Results

Serum(-):

Slide 500: Quotion picture, Tet serum (-), 1:10 diluted
Slide 500: binding curves

Serum(+):

Slide 024: Quotion picture, Tet serum (+), 1:10 diluted

→ bad measurement, no binding curves were obtained

Slide 504: Quotion picture, Tet serum (+), 1:30 diluted
Slide 504: binding curves

→ signal for 1:10 dilution of the blood serum much better → continue with 1:10 dilution

Experiment 64b: Tetanus antigen PDITC slides for iRIf III

2015.08.31

Experiment/Protocol

  • 4 PDITC slides were prepared
  • Plasma activation: 40 L/h, 5 min
  • APTES incubation: 30 min
  • PDITC incubation: 2 h
  • desiccator: 15 min
  • stored at 4 °C

2015.09.02

Experiment/Protocol

Spotting pattern:

# spot Concentration
1 pos. control (GFP) 0.5 mg/ml
2 mCherry 0.5 mg/ml
3 Tetanus Antigen ~0.5 mg/ml
4 BSA 10 mg/ml
5 BSA +bBSA 10 mg/ml
6 bBSA 200 µg/ml

note: bBSA on spot 5 was incubated for only a minute and then replaced with BSA

2015.09.03

Experiment/Protocol

Flush protocol:

Tet serum (-): 287 (1:10 diluted serum),429 (1:30 diluted serum)
Tet serum (+): 443 (1:10 diluted serum), 423 (1:30 diluted serum)

Reagent Step Flow rate (ul/min) Priming time Concentration
Buffer1606001x
BSA26060010 mg/ml
Buffer3606001x
anti-GFP4306001.5 ug/ml
Buffer5603001x
Serum SIG002 (-)/(+)6209001:10
Buffer7603001x
StrepCy58306005 ug/ml
Buffer9603001x

Results

Serum(-):

Slide 287: Quotion picture, Tet serum (-), 1:10 diluted
Slide 287: binding curves
Slide 429: Quotion picture, Tet serum (-), 1:30 diluted
Slide 429: binding curves

→ higher dilution of the blood serum leads to weaker iRif signal for mCherry, BSA,BSA+bBSA and bBSA but also for the tetanus antigen

Serum(+):

Slide 443: Quotion picture, Tet serum (+), 1:10 diluted
Slide 443: binding curves
Slide 423: Quotion picture, Tet serum (+), 1:30 diluted
Slide 423: binding curves

→ higher dilution of the blood serum leads to weaker iRif signal for mCherry, BSA,BSA+bBSA and bBSA but also for the tetanus antigen

→ signal of the tetanus antigen spot very weak compared to experiment 59a → use fresh stock of tetanus antigen next time

→ unspecific binding to the BSA spot is most of the time weaker than to mCherry → from now on BSA will be used as negative contol for blood serum

Experiment 64a: Salmonella antigen on PDITC slides for iRIf II

2015.08.31

Experiment/Protocol

  • 3 PDITC slides were prepared
  • Plasma activation: 40 L/h, 5 min
  • APTES incubation: 30 min
  • PDITC incubation: 2 h
  • desiccator: 15 min
  • stored at 4 °C

2015.09.01

Experiment/Protocol

Spotting pattern:

# spot Concentration
1 pos. control (GFP) 0.5 mg/ml
2 bBSA 0.5 mg/ml
3 Salmonella Antigen (15) undiluted

Slides (3): 208,215,216

2015.09.02

Experiment/Protocol

Flush protocol:

Reagent Step Flow rate (ul/min) Priming time Concentration
Buffer (TBS)1606001x
BSA26060010 mg/ml
Buffer (TBS)3606001x
anti-GFP4306001,5 ug/ml
Buffer (TBS)5603001x
anti-Salmonella (13, desalt)630900undiluted
Buffer (TBS)7603001x
StrepCy58306005 ug/ml
Buffer (TBS)9603001x

last slide (208) changed protocol: only 1 ug/ml anti-GFP used; anti-Salmonella desalt was mixed 1:1 with anti-Salmonella lysate

Results

Note: Slide 208 had the best signal.

Slide 208: ROI selection
Slide 208: Binding curves