Protocol: Silver Staining
Preparation of Gel
Ingredients
Note: Please, adapt the final percentage of polyacrilamide according to the size of the protein.
Expected protein size | Final polyacrilamide concentration |
4-40 kDA | 20% |
12-45 kDA | 15% |
10-70 kDA | 12.5% |
15-100 kDA | 10% |
25-200 kDA | 8% |
For a Gel percentage of 7.5 % the proportions are the following:
Separating gel | Volume, en mL |
30% Polyacrylamide | 10 |
1.5M Tris (pH 8.8) | 10 |
10% Ammonium persulfate | 0.4 |
10% SDS | 0.4 |
TEMED | 0.038 |
H2O | 19.2 |
Total volume | 40 |
Stacking gel | Volume, en mL |
30% Polyacrylamide | 1.36 |
1M Tris (pH 6.8) | 1 mL |
10% Ammonium persulfate | 0.08 |
10% SDS | 0.08 |
TEMED | 0.008 |
H2O | 5.44 |
Total volume | 8 mL |