Smith Lab

Silver Staining

  1. Rinse gel plates and staining glass dish with 50% nitric acid to remove everything.
  2. After electrophoresis, fix gels (2 minigels = ~200 ml, 2 large gels = ~400 ml) in 50% MeOH/0.1% Acetic Acid.  20 min-1hour
  3. Wash gels in dH2O.  15 min.
  4. Soak gels in 0.001% DTT.  30 min.
  5. Soak gels in 0.1% Silver Nitrate. 30 min.
  6. Rinse gels with dH20 to remove excess silver.  3 X 10 sec.
  7. Develop in fresh 3% Sodium Carbonate and Formaldehyde(50 ul of 37%/100 ml). 
  8. Stop reaction with 2 M Citric Acid and incubate ~10 min to make sure reaction is quenched.  (~10 ml/100ml solution)
  9. Soak gel in dH2O 10 min.