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Star Republic: Guide for Biologists

Silver stain of proteins

1. Wear gloves, rinse gloves and glasswares with ddH2O.

2. Soak the polyamcriamide gel in 50% methanol (> 1 hour).

3. Change to fresh 50% methanol and leave for 1 hour.

4. Make fresh stain solution. Add 1.4 ml Ammonium hydroxide (use the fresh, frozen stock aliquot in the -20 C freezer), 0.185 ml 10 M NaOH, 4.0 ml 20% (w/v) AgNO3 (add dropwise), and ddH2O to 100 ml.

5. Rinse the gel with ddH2O.

6. Add the staining solution. Agitate for 10 minutes at room temperature.

7. Discard the staining solution and rinse the gel with ddH2O.

8. Soak for 5 minutes in fresh ddH2O.

9. Rinse again briefly in ddH2O.

10. Add developer solution (0.01% (w/v) citric acid (1% w/v), 0.038% formaldehyde) and agitate for 2 to 10 minutes.

11. When the gel achieves the desired state of staining, stop with 50% methanol/10% acetic acid.

12. Store the gel in 50% methanol/10% acetic acid.