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Non-Destructive Silver Staining (modified for MS)
Fix the gel for at least an hour by gently shaking in 40% Ethanol [EtOH]/10% Acetic Acid [HAc]/50% Millipore water (200ml or about five times gel volume).
Washing and Sensitizing
Wash the gel two (2) times for 20 minutes each in 30% EtOH (same volume requirements as above).
Wash the gel once with Millipore water for 20 minutes
Sensitize the gel in 0.02% Sodium Thiosulfate [Na
] for 1 minute.
Wash the gel three (3) times for 30 seconds with Millipore water.
Staining and Developing
Incubate the gel for 20 minutes with cold 0.1% Silver Nitrate [AgNO
]/0.02% formal [37% formaldehyde] solution.
Wash the gel four (4) times for 30 seconds with Millipore water (room temperature).
Develop the gel in 3% Sodium Carbonate [Na
]/0.05% formalin, observe the color and change solution when developer turns yellow. Stop the reaction when staining is sufficient.
Terminating Staining and Storage
Wash the gel with Millipore water for 30 seconds.
Terminate development by incubating in 5% HAc for 5 minutes.
Wash the gel three (3) times for 10 minutes with Millipore water.
If gel is to be stored for longer time, keep immersed in 1% HAc at 4C.
Blum et al.