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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 | |
| Gel percentage | 5 % |
| 30% Polyacrylamide | 1.36 mL |
| 1M Tris (pH 6.8) | 1 mL |
| 10% Ammonium persulfate | 0.08 mL |
| 10% SDS | 0.08 mL |
| TEMED | 0.008 mL |
| H2O | 5.44 mL |
| Total volume | 8 mL |
