- 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 [Na2S2O3] 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 [AgNO3]/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 [Na2CO3]/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. Electrophoresis. 1987; 8: 93-99.