Team:UiOslo Norway/Experiments/SDS-Page

SDS-Page:

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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