Silver Development protocol (silver stain) for 0.75mm polyacrylamide gels
Agitate gel twice with 50% MeOH, 10% acetic acid (Fixer 1) for 15 min ea.
Rinse (1 min) with 200 ml DW.
Agitate in 10% EtOH, 5% glacial acetic acid (Fixer 2) for 6 min.
Agitate in distilled water.
Agitate twice (9 min each) with 500 ml DW.
Agitate in 500 ml of 20 mg/L Na2S2O4 (hydrosulfite [dithionite], make fresh) in distilled, deionized water (DDW) for 9min (sensitization).
Pour off solution.
Agitate for 9 min.
Rinse 30 sec with DDW to remove excess AgNO3.
Add 200 ml image developer (1ml 37% formaldehyde per liter of 3% sodium carbonate) mixed with 200 ml 10g/L sodium thiosulfate.
Pour off developer.
Add 80 ml of stop solution (50 g tris, 25 ml glacial acetic per liter of distilled water).
Store gel in 10% glycerol with bacteriostat (0.02% NaN3 ).
Without rinsing, add 200 ml, 0.1% AgNO3 (200 mg) in DDW.
Add 150 ml 37% formaldehyde.
Agitate to desired staining intensity (3 to 6 min).
