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

Line 45: Line 45:
 
     <td>4.8 ml    </td>
 
     <td>4.8 ml    </td>
 
     <td>30 % Acrylamide (37.5:1)</td>
 
     <td>30 % Acrylamide (37.5:1)</td>
 +
    </tr>
 +
  <tr>
 +
    <td>2.7 ml </td>
 +
    <td>1.1 ml   </td>
 +
    <td>Water</td>
 +
    </tr>
 +
  <tr>
 +
    <td>80 μl </td>
 +
    <td>80 μl  </td>
 +
    <td>10% (w/v) SDS</td>
 +
    </tr>
 +
  <tr>
 +
    <td>8 μl</td>
 +
    <td>8 μl </td>
 +
    <td>TEMED</td>
 +
    </tr>
 +
  <tr>
 +
    <td>80 μl </td>
 +
    <td>80 μl </td>
 +
    <td>APS</td>
 
     </tr>
 
     </tr>
  

Revision as of 11:57, 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)
    2.7 ml 1.1 ml Water
    80 μl 80 μl 10% (w/v) SDS
    8 μl 8 μl TEMED
    80 μl 80 μl APS

    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