Difference between revisions of "Team:UiOslo Norway/Experiments/SDS-Page"

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   <li><p><b>Separating gel:</b></br>
 
   <li><p><b>Separating gel:</b></br>
 
</br>
 
</br>
−
12 % 18 %</br>
+
<table id="t01">
−
<hr style="height:2px;border:none;color:#472400;background-color:#333;" />
+
  <tr>
−
<p>2 ml 2 ml Seperating buffer (1.5 M Tris pH 8.8)</br>
+
    <th>12 % </th>
−
3.2 ml 4.8 ml   30 % Acrylamide (37.5:1)</br>
+
    <th>18 % </th>
−
2.7 ml 1.1 ml Water</br>
+
  </tr>
−
80 ul 80 ul 10% (w/v) SDS</br>
+
  <tr>
−
8 ul 8 ul TEMED</br>
+
    <td>2 ml</td>
−
80 ul 80 ul APS</br>
+
    <td>2 ml</td>
 +
    <td>Seperating buffer (1.5 M Tris pH 8.8)</td>
 +
  </tr>
 +
  <tr>
 +
    <td>3.2 ml </td>
 +
    <td>4.8 ml   </td>
 +
    <td>30 % Acrylamide (37.5:1)</td>
 +
    </tr>
 +
 
 +
</p></table>
 +
 
 
</br>
 
</br>
−
Run SDS-Page for 1 hour at 30 mA per gel
+
<p>Run SDS-Page for 1 hour at 30 mA per gel</p>
 
</li>
 
</li>
 
</ol>
 
</ol>

Revision as of 11:55, 14 August 2015

SDS-Page:

Back to Protocols


Reagent


  1. Stacking Gel:

    0.7 ml 30 % Acrylamide (37.5:1)
    1.25 ml Stacking gel buffer (1 M Tris pH 6.8)
    3 ml Water
    50 ul 10 % (w/v) SDS
    5 ul TEMED
    50 ul APS


  2. Separating gel:

    12 % 18 %
    2 ml 2 ml Seperating buffer (1.5 M Tris pH 8.8)
    3.2 ml 4.8 ml 30 % Acrylamide (37.5:1)

    Run SDS-Page for 1 hour at 30 mA per gel

Coomassie-staining

Quick Coomassie staining solution:

1 % (w/v) Coomassie Brilliant Blue R250
50 % (v/v) Methanol

10 % (v/v) Acetic acid
Coomassie Destaining solution:
40 % (v/v) Methanol

10 % (v/v) Acetic acid