Silver staining

Solutions

  • 50% methanol/water
  • 1 g/L AgNO3
  • 30 g/L Na2CO3
  • 1 M DTT
  • 2 M citric acid/water

Procedure

  • Fix gel by shaking in 50% methanol 2× 15 min
  • Rehydrate in water + DTT (50 mL + 5 µL 1 M DTT) 15 min
  • Shake in 0.1% AgNO3 (50 mL) 20 min
  • Prepare fresh 50 mL 3% Na2CO3 + 100 µL formaldehyde (30%)
  • Rinse gel with 10 mL of above solution until it turns brown, discard
  • Add remainder of solution and shake until bands are visible
  • Stop by adding 2 M citric acid solution
  • Rinse with water 2–3× 5 min

Adapted from the Salk Institute Mass Spectrometry Core Facility (with permission)
Original document at http://cores.salk.edu/massspec/protocols/silver.php