Siver Stain Protocol
(from Dunbar, B.S. 1987. Two-Dimesional Electrophoresis and Immunological Techniques. Plenum Press, NY. 372 pp)
- 0.8 g silver nitrate
- Bring to 4 mL in distilled water
- 21 mL 0.36% NaOH
- 1.4 mL 14.8 M ammonium hydroxide
- For best results, make fresh daily
- add Solution A dropwise to B with constant stirring then bring to 100 mL with distilled water.
- NOTE: SOLUTION C MUST BE USED WITHIN 5 MIN
- 2.5 mL 1% citric acid
- 0.25 mL 37% formaldehyde
- Make to 500 mL with distilled water.
- SOLUTION D MUST BE “VERY FRESH”
- #1: 50 % methanol
- #2: 5% acetic acid
- Soak gel in 50% MeOH at least 60 min
- Stain gel in Solution C (15 min with gentle agitation)
- Wash gel in water (5 min with gentle agitation)
- Develop gel in Solution D
- Stop with 5% acetic acid
- Wash gel with water and place in 50% MeOH overnight.