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

 
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<h1 class="sectionedit1">Labjournal iRIf September</h1>
 
<h1 class="sectionedit1">Labjournal iRIf September</h1>
 
<div class="level1">
 
<div class="level1">
 
</div>
 
</div>
−
<!-- EDIT1 SECTION "Labjournal iRIf September" [1-42] -->
+
<!-- EDIT1 SECTION "Labjournal iRIf September" [1-40] -->
−
<h3 class="sectionedit2">Measuring own blood: Tetanus (-), Tetanus (+)</h3>
+
<h3 class="sectionedit2">Measuring own blood (tetanus vaccination)</h3>
 +
<div class="level3">
 +
<p>
 +
<strong>16.09.2015 (spotted)</strong>
 +
</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/>
 +
</p>
 +
<div class="table sectionedit3"><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>
 +
<!-- EDIT3 TABLE [200-511] -->
 +
<p>
 +
<strong>17.09.2015 (measured)</strong>
 +
</p>
 +
<p>
 +
Tet serum (-+): 024, 466 <br/>
 +
</p>
 +
<div class="table sectionedit4"><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>
 +
<!-- EDIT4 TABLE [568-848] -->
 +
<p>
 +
Tet serum (+-): 208, 216 <br/>
 +
</p>
 +
<div class="table sectionedit5"><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>
 +
<!-- EDIT5 TABLE [880-1160] -->
 +
<p>
 +
GFP; Tet serum (-+): 287 <br/>
 +
</p>
 +
<div class="table sectionedit6"><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>
 +
<!-- EDIT6 TABLE [1192-1472] -->
 +
</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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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>
 +
<!-- EDIT2 SECTION "Measuring own blood (tetanus vaccination)" [41-2900] -->
 +
<h3 class="sectionedit7">Measuring : The **DIA Chip**</h3>
 +
<div class="level3">
 +
<p>
 +
<strong>14.09.2015 (stamped)</strong>
 +
</p>
 +
<p>
 +
PDMS Dia-Chip stamp (incubated 30 minutes with 1 mg/ml GFP)
 +
was stamped onto plasmaactivated slide.
 +
Kept in 10 mg/ml BSA in PBS at 4° until measurement.
 +
</p>
 +
<p>
 +
<strong>16.09.2015 (measured)</strong>
 +
</p>
 +
<p>
 +
Slides were previously blocked with the backside of a PDMS flowcell to avoid
 +
airbubble issues due to the hydrophobicity of the stamp.
 +
</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">anti-GFP</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">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</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 [3284-3518] --><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/a/a5/Freiburg_labjournal-irif-diachip.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:irif:diachip.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 287: Cy5 signal after iRIf measurement detected in microarray scanner</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-irif-diachipirif.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:irif:diachipirif.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 303: Quotient picture of the anti-GFP step</div></div></div>
 +
</div>
 +
<!-- EDIT7 SECTION "Measuring : The **DIA Chip**" [2901-3743] -->
 +
<h3 class="sectionedit9">Rabbit-serum (GFP immunized/not immunized) on PDITC</h3>
 +
<div class="level3">
 +
<p>
 +
<strong>14.09.2015 (spotted)</strong>
 +
</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/>
 +
</p>
 +
<div class="table sectionedit10"><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 (bBSA) </td><td class="col2"> 0.2 mg/ml </td>
 +
</tr>
 +
<tr class="row2">
 +
<td class="col0"> 2 </td><td class="col1"> neg. control (BSA)</td><td class="col2"> 0.2 mg/ml </td>
 +
</tr>
 +
<tr class="row3">
 +
<td class="col0"> 3 </td><td class="col1"> His-GFP desalt </td><td class="col2"> 1mg/ml </td>
 +
</tr>
 +
<tr class="row4">
 +
<td class="col0"> 4 </td><td class="col1"> pos. control (bBSA) </td><td class="col2"> 0.2 mg/ml </td>
 +
</tr>
 +
<tr class="row5">
 +
<td class="col0"> 5 </td><td class="col1"> neg. control (BSA)</td><td class="col2"> 0.2 mg/ml</td>
 +
</tr>
 +
<tr class="row6">
 +
<td class="col0"> 6 </td><td class="col1"> His-GFP desalt </td><td class="col2"> 1mg/ml </td>
 +
</tr>
 +
<tr class="row7">
 +
<td class="col0"> 7 </td><td class="col1">His-mCherry desalt</td><td class="col2"> 1 mg/ml </td>
 +
</tr>
 +
<tr class="row8">
 +
<td class="col0"> 8 </td><td class="col1">His-mCherry desalt</td><td class="col2"> 1 mg/ml </td>
 +
</tr>
 +
</table></div>
 +
<!-- EDIT10 TABLE [3912-4235] -->
 +
<p>
 +
<strong>15.09.2015 (measured)</strong>
 +
</p>
 +
<p>
 +
Flush protocol:
 +
Positive serum (504,424,466).
 +
Negative serum (423).
 +
</p>
 +
<div class="table sectionedit11"><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">StrepCy5</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">60</td><td class="col3">300</td><td class="col4">1x</td>
 +
</tr>
 +
<tr class="row6">
 +
<td class="col0">Serum (+)/(-)</td><td class="col1">8</td><td class="col2">30</td><td class="col3">900</td><td class="col4">1:30</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">Anti-Rabbit</td><td class="col1">10</td><td class="col2">60</td><td class="col3">300</td><td class="col4">5 ug/ml</td>
 +
</tr>
 +
<tr class="row9">
 +
<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>
 +
<!-- EDIT11 TABLE [4335-4625] -->
 +
<p>
 +
*Slide 466 was measured with 10 ug/ml Anti-Rabbit
 +
</p>
 +
<p>
 +
Results: Serum (-) fromm rabbit that was not immunized to GFP shows no binding to the GFP spot.<br/>
 +
 
 +
Serum (+) from rabbit that was immunized to GFP shows extensive binding to the GFP spot.<br/>
 +
</p>
 +
<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-irif-roi_selection_20150915_423_cropped.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:irif:roi_selection_20150915_423_cropped.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 423 (serum(-)): Selections 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/b/b7/Freiburg_labjournal-irif-binding_curve_20150915_423.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:irif:binding_curve_20150915_423.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 423 (serum(-)): 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/2015/5/5e/Freiburg_labjournal-irif-roi_selection_20150915_424_cropped.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:irif:roi_selection_20150915_424_cropped.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 424 (serum(+)): 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/b/b1/Freiburg_labjournal-irif-binding_curves_20150915_424.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:irif:binding_curves_20150915_424.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 424 (serum(+)): Binding curves</div></div></div><hr/>
 +
<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><a class="media" href="https://static.igem.org/mediawiki/2015/8/8e/Freiburg_labjournal-irif-roi_selection_20150915_466_cropped.png" title="labjournal:irif:roi_selection_20150915_466_cropped.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/8/8e/Freiburg_labjournal-irif-roi_selection_20150915_466_cropped.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/8/8e/Freiburg_labjournal-irif-roi_selection_20150915_466_cropped.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 466 (serum(+)): Selections of ROIs</div></div></div>
 +
</div>
 +
<!-- EDIT9 SECTION "Rabbit-serum (GFP immunized/not immunized) on PDITC" [3744-5418] -->
 +
<h3 class="sectionedit12">Measuring own blood (Tetanus vaccination)</h3>
 +
<div class="level3">
 +
<p>
 +
<strong>13.09.2015 (spotted)</strong>
 +
</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/>
 +
</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 [5577-5760] -->
 +
<p>
 +
<strong>14.09.2015 (measured)</strong>
 +
</p>
 +
<p>
 +
Tet serum (-): 208, 254 <br/>
 +
 
 +
Tet serum (+): 500, 024
 +
</p>
 +
<div class="table sectionedit14"><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>
 +
<!-- EDIT14 TABLE [5841-6188] -->
 +
</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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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>
 +
<!-- EDIT12 SECTION "Measuring own blood (Tetanus vaccination)" [5419-7134] -->
 +
<h3 class="sectionedit15">Measuring own blood serum (Tetanus vaccination)</h3>
 +
<div class="level3">
 +
<p>
 +
<strong>12.09.2015 (spotted)</strong>
 +
</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/>
 +
</p>
 +
<div class="table sectionedit16"><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>
 +
<!-- EDIT16 TABLE [7299-7482] -->
 +
<p>
 +
<strong>13.09.2015 (measured)</strong>
 +
</p>
 +
<p>
 +
Tet serum (-): 443, 466 <br/>
 +
 
 +
Tet serum (+): 423, 424
 +
</p>
 +
<div class="table sectionedit17"><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>
 +
<!-- EDIT17 TABLE [7563-7910] -->
 +
</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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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>
 +
<!-- EDIT15 SECTION "Measuring own blood serum (Tetanus vaccination)" [7135-8854] -->
 +
<h3 class="sectionedit18">Measuring own blood (Tetanus vaccination)</h3>
 +
<div class="level3">
 +
<p>
 +
<strong>05.09.2015 (spotted)</strong>
 +
</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/>
 +
</p>
 +
<div class="table sectionedit19"><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>
 +
<!-- EDIT19 TABLE [9012-9195] -->
 +
<p>
 +
<strong>06.09.2015 (measured)</strong>
 +
</p>
 +
<p>
 +
Flush protocol:
 +
</p>
 +
<p>
 +
Tet serum (-): 443 <br/>
 +
 
 +
Tet serum (+): 466
 +
</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">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>
 +
<!-- EDIT20 TABLE [9284-9577] -->
 +
<p>
 +
Tet serum (-): 208<br/>
 +
 
 +
Tet serum (+): 303
 +
</p>
 +
<div class="table sectionedit21"><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>
 +
<!-- EDIT21 TABLE [9621-9914] -->
 +
<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%3Airif%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%3Airif%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><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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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>
 +
<!-- EDIT18 SECTION "Measuring own blood (Tetanus vaccination)" [8855-11182] -->
 +
<h3 class="sectionedit22">Measuring own blood: Tetanus (-), Tetanus (+)</h3>
 
<div class="level3">
 
<div class="level3">
 
<p>
 
<p>
Line 28: Line 678:
 
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 sectionedit3"><table class="inline">
+
<div class="table sectionedit23"><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 51: Line 701:
 
</tr>
 
</tr>
 
</table></div>
 
</table></div>
−
<!-- EDIT3 TABLE [204-387] -->
+
<!-- EDIT23 TABLE [11344-11527] -->
 
<p>
 
<p>
 
Flush protocol:
 
Flush protocol:
 
</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 sectionedit4"><table class="inline">
+
<div class="table sectionedit24"><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 80: Line 730:
 
</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">1:10/1:30</td>
 
</tr>
 
</tr>
 
<tr class="row7">
 
<tr class="row7">
Line 92: Line 742:
 
</tr>
 
</tr>
 
</table></div>
 
</table></div>
−
<!-- EDIT4 TABLE [441-740] -->
+
<!-- EDIT24 TABLE [11630-11920] -->
 
</div>
 
</div>
−
<!-- EDIT2 SECTION "Measuring own blood: Tetanus (-), Tetanus (+)" [43-740] -->
+
<h4>Results</h4>
−
<h3 class="sectionedit5">Specific surface (Ni-NTA) vs. unspecific surface (PDITC): cellfree expressed GFP</h3>
+
<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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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>
 +
<!-- EDIT22 SECTION "Measuring own blood: Tetanus (-), Tetanus (+)" [11183-12903] -->
 +
<h3 class="sectionedit25">Specific surface (Ni-NTA) vs. unspecific surface (PDITC): cellfree expressed GFP</h3>
 
<div class="level3">
 
<div class="level3">
 
<p>
 
<p>
Line 103: Line 778:
 
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 sectionedit6"><table class="inline">
+
<div class="table sectionedit26"><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>
−
<!-- EDIT6 TABLE [938-1295] -->
+
<!-- EDIT26 TABLE [13101-13458] -->
 
<p>
 
<p>
 
<strong>05.09.2015 (measured)</strong>
 
<strong>05.09.2015 (measured)</strong>
Line 139: Line 814:
 
Flush protocol:
 
Flush protocol:
 
</p>
 
</p>
−
<div class="table sectionedit7"><table class="inline">
+
<div class="table sectionedit27"><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>
−
<!-- EDIT7 TABLE [1342-1578] -->
+
<!-- EDIT27 TABLE [13505-13741] -->
 +
<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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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%3Airif%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>
−
<!-- EDIT5 SECTION "Specific surface (Ni-NTA) vs. unspecific surface (PDITC): cellfree expressed GFP" [741-1581] -->
+
<!-- EDIT25 SECTION "Specific surface (Ni-NTA) vs. unspecific surface (PDITC): cellfree expressed GFP" [12904-14826] -->
−
<h3 class="sectionedit8">Measuring own blood: Tetanus (-), Tetanus (+) Sig002</h3>
+
<h3 class="sectionedit28">Measuring own blood: Tetanus (-), Tetanus (+) Sig002</h3>
 
<div class="level3">
 
<div class="level3">
 
<p>
 
<p>
Line 174: Line 863:
 
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 sectionedit9"><table class="inline">
+
<div class="table sectionedit29"><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>
−
<!-- EDIT9 TABLE [1750-1966] -->
+
<!-- EDIT29 TABLE [14995-15211] -->
 
<p>
 
<p>
 
<strong>03.09.2015 (measured)</strong>
 
<strong>03.09.2015 (measured)</strong>
Line 205: Line 894:
 
Tet (+): 443, 423 (1:30 diluted serum)
 
Tet (+): 443, 423 (1:30 diluted serum)
 
</p>
 
</p>
−
<div class="table sectionedit10"><table class="inline">
+
<div class="table sectionedit30"><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>
−
<!-- EDIT10 TABLE [2073-2367] --><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/75/Freiburg_labjournal-roi_selection_20150903_287_-1_10_.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:roi_selection_20150903_287_-1_10_.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 287: ROI selection, ROIs were chosen in similar manner for the other slides</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/ec/Freiburg_labjournal-binding_curves_20150903_287_-_1_10_.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:binding_curves_20150903_287_-_1_10_.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 287: Binding curves; serum (-) diluted 1:10</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/20/Freiburg_labjournal-binding_curves_20150903_443_1_10_.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:binding_curves_20150903_443_1_10_.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 443: Binding curves; serum (+) diluted 1:10</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/7c/Freiburg_labjournal-binding_curves_20150903_429_-_1_30_.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:binding_curves_20150903_429_-_1_30_.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 429: Binding curves; serum (-) diluted 1:30</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/6c/Freiburg_labjournal-binding_curves_20150903_423_1_30_.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:binding_curves_20150903_423_1_30_.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 423: Binding curves, serum (+) diluted 1:30</div></div></div>
+
<!-- EDIT30 TABLE [15318-15612] --><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/75/Freiburg_labjournal-roi_selection_20150903_287_-1_10_.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:roi_selection_20150903_287_-1_10_.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 287: ROI selection, ROIs were chosen in similar manner for the other slides</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/ec/Freiburg_labjournal-binding_curves_20150903_287_-_1_10_.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:binding_curves_20150903_287_-_1_10_.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 287: Binding curves; serum (-) diluted 1:10</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/20/Freiburg_labjournal-binding_curves_20150903_443_1_10_.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:binding_curves_20150903_443_1_10_.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 443: Binding curves; serum (+) diluted 1:10</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/7/7c/Freiburg_labjournal-binding_curves_20150903_429_-_1_30_.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:binding_curves_20150903_429_-_1_30_.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 429: Binding curves; serum (-) diluted 1:30</div></div></div><div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/6c/Freiburg_labjournal-binding_curves_20150903_423_1_30_.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:binding_curves_20150903_423_1_30_.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div>Slide 423: Binding curves, serum (+) diluted 1:30</div></div></div>
 
</div>
 
</div>
−
<!-- EDIT8 SECTION "Measuring own blood: Tetanus (-), Tetanus (+) Sig002" [1582-2990] -->
+
<!-- EDIT28 SECTION "Measuring own blood: Tetanus (-), Tetanus (+) Sig002" [14827-16235] -->
−
<h3 class="sectionedit11">Distinguish between different antigen/antibody interaction (Salmonella) (on PDITC) [finished]</h3>
+
<h3 class="sectionedit31">Distinguish between different antigen/antibody interaction (Salmonella) (on PDITC) [finished]</h3>
 
<div class="level3">
 
<div class="level3">
 
<p>
 
<p>
Line 248: Line 937:
 
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/>
 
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/>
 
</p>
 
</p>
−
<div class="table sectionedit12"><table class="inline">
+
<div class="table sectionedit32"><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>
−
<!-- EDIT12 TABLE [3202-3342] -->
+
<!-- EDIT32 TABLE [16447-16587] -->
 
<p>
 
<p>
 
Slides (3): 208,215,216
 
Slides (3): 208,215,216
Line 273: Line 962:
 
Flush protocol:
 
Flush protocol:
 
</p>
 
</p>
−
<div class="table sectionedit13"><table class="inline">
+
<div class="table sectionedit33"><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 [3541-3878] -->
+
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<div class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/8/89/Freiburg_labjournal-roi_selection_20150902_208.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:roi_selection_20150902_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/b/bc/Freiburg_labjournal-binding_curve_20150902_208.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:binding_curve_20150902_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"><a class="media" href="https://static.igem.org/mediawiki/2015/0/02/Freiburg_labjournal-binding_curves_20150902_215.png" title="labjournal:binding_curves_20150902_215.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/0/02/Freiburg_labjournal-binding_curves_20150902_215.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/0/02/Freiburg_labjournal-binding_curves_20150902_215.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"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/9/91/Freiburg_labjournal-binding_curves_20150902_216.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:binding_curves_20150902_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 class="thumb2 trien" style="width:210px"><div class="thumbinner"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/8/89/Freiburg_labjournal-roi_selection_20150902_208.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:roi_selection_20150902_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/b/bc/Freiburg_labjournal-binding_curve_20150902_208.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:binding_curve_20150902_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"><a class="media" href="https://static.igem.org/mediawiki/2015/0/02/Freiburg_labjournal-binding_curves_20150902_215.png" title="labjournal:binding_curves_20150902_215.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/0/02/Freiburg_labjournal-binding_curves_20150902_215.png" width="200"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/0/02/Freiburg_labjournal-binding_curves_20150902_215.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"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/9/91/Freiburg_labjournal-binding_curves_20150902_216.png" width="200"/><div class="thumbcaption"><div class="magnify"><a class="internal" href="/igem2015/lib/exe/detail.php?id=labjournal%3Airif%3Aseptember&amp;media=labjournal:binding_curves_20150902_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>
 
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Latest revision as of 07:14, 20 November 2015

""

<< Back

Labjournal iRIf September

Measuring own blood (tetanus vaccination)

16.09.2015 (spotted)

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

17.09.2015 (measured)

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

Measuring : The **DIA Chip**

14.09.2015 (stamped)

PDMS Dia-Chip stamp (incubated 30 minutes with 1 mg/ml GFP) was stamped onto plasmaactivated slide. Kept in 10 mg/ml BSA in PBS at 4° until measurement.

16.09.2015 (measured)

Slides were previously blocked with the backside of a PDMS flowcell to avoid airbubble issues due to the hydrophobicity of the stamp.

Reagent Step Flow rate (ul/min) Priming time Concentration
Buffer1603001x
BSA26060010 mg/ml
Buffer3606001x
anti-GFP4306005 ug/ml
Buffer5603001x
StrepCy56603005 ug/ml
Buffer9603001x
Slide 287: Cy5 signal after iRIf measurement detected in microarray scanner
Slide 303: Quotient picture of the anti-GFP step

Rabbit-serum (GFP immunized/not immunized) on PDITC

14.09.2015 (spotted)

Spotting pattern:

# spot Concentration
1 pos. control (bBSA) 0.2 mg/ml
2 neg. control (BSA) 0.2 mg/ml
3 His-GFP desalt 1mg/ml
4 pos. control (bBSA) 0.2 mg/ml
5 neg. control (BSA) 0.2 mg/ml
6 His-GFP desalt 1mg/ml
7 His-mCherry desalt 1 mg/ml
8 His-mCherry desalt 1 mg/ml

15.09.2015 (measured)

Flush protocol: Positive serum (504,424,466). Negative serum (423).

Reagent Step Flow rate (ul/min) Priming time Concentration
Buffer1603001x
BSA26060010 mg/ml
Buffer3606001x
StrepCy54306005 ug/ml
Buffer5603001x
Serum (+)/(-)8309001:30
Buffer9606001x
Anti-Rabbit10603005 ug/ml
Buffer11603001x

*Slide 466 was measured with 10 ug/ml Anti-Rabbit

Results: Serum (-) fromm rabbit that was not immunized to GFP shows no binding to the GFP spot.
Serum (+) from rabbit that was immunized to GFP shows extensive binding to the GFP spot.

Slide 423 (serum(-)): Selections of ROIs
Slide 423 (serum(-)): Binding curves
Slide 424 (serum(+)): Selection of ROIs
Slide 424 (serum(+)): Binding curves
Slide 466 (serum(+)): Selections of ROIs

Measuring own blood (Tetanus vaccination)

13.09.2015 (spotted)

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

14.09.2015 (measured)

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

Measuring own blood serum (Tetanus vaccination)

12.09.2015 (spotted)

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

13.09.2015 (measured)

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

Measuring own blood (Tetanus vaccination)

05.09.2015 (spotted)

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

06.09.2015 (measured)

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

Measuring own blood: Tetanus (-), Tetanus (+)

04.09.15 (spotted)

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

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 serum6306001:10/1:30
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

Specific surface (Ni-NTA) vs. unspecific surface (PDITC): cellfree expressed GFP

04.09.2015 (spotted)

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

05.09.2015 (measured)

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

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

Measuring own blood: Tetanus (-), Tetanus (+) Sig002

02.09.2015 (spotted) 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

03.09.2015 (measured)

Tet (-): 287,429 (1:30 diluted serum) Tet (+): 443, 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
Slide 287: ROI selection, ROIs were chosen in similar manner for the other slides
Slide 287: Binding curves; serum (-) diluted 1:10
Slide 443: Binding curves; serum (+) diluted 1:10
Slide 429: Binding curves; serum (-) diluted 1:30
Slide 423: Binding curves, serum (+) diluted 1:30

Distinguish between different antigen/antibody interaction (Salmonella) (on PDITC) [finished]

01.09.2015 (spotted)

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

02.09.2015 (measured)

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

Note: Background of Slides 208 looked nicest.

Slide 208: ROI selection
Slide 208: Binding curves
Slide 215: Binding curves
Slide 216: Binding curves