Siver Stain Protocol

(from Dunbar, B.S. 1987. Two-Dimesional Electrophoresis and Immunological Techniques. Plenum Press, NY. 372 pp)

Solutions

 Solution A:

 0.8 g silver nitrate

Bring to 4 mL in distilled water
 Solution B:

21 mL 0.36% NaOH

1.4 mL 14.8 M ammonium hydroxide

For best results, make fresh daily
 Solution C: 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
 Solution D:

2.5 mL 1% citric acid

0.25 mL 37% formaldehyde

Make to 500 mL with distilled water.

SOLUTION D MUST BE "VERY FRESH"
Misc Solutions:

#1: 50 % methanol

#2: 5% acetic acid

Protocol

1. Soak gel in 50% MeOH at least 60 min

2. Stain gel in Solution C (15 min with gentle agitation)

3. Wash gel in water (5 min with gentle agitation)

4. Develop gel in Solution D

5. Stop with 5% acetic acid

6. Wash gel with water and place in 50% MeOH overnight.