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

Line 45: Line 45:
 
<p>
 
<p>
 
<p><h3>Coomassie-staining </h3>
 
<p><h3>Coomassie-staining </h3>
<p>Quick Coomassie staining solution:
</br>
+
<p><b>Quick Coomassie staining solution:
</b></br>
 
1 % (w/v) Coomassie Brilliant Blue R250</br>  
 
1 % (w/v) Coomassie Brilliant Blue R250</br>  
 
50 % (v/v) Methanol
</br>
 
50 % (v/v) Methanol
</br>
 
10 % (v/v) Acetic acid </br>
 
10 % (v/v) Acetic acid </br>
Coomassie Destaining solution: </br>
+
<b>Coomassie Destaining solution:</b> </br>
 
40 % (v/v) Methanol
</br>
 
40 % (v/v) Methanol
</br>
 
10 % (v/v) Acetic acid </br>
 
10 % (v/v) Acetic acid </br>

Revision as of 11:49, 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 ul 80 ul 10% (w/v) SDS
    8 ul 8 ul TEMED
    80 ul 80 ul 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