CCs Magical Silver Stain

1. Fix gel in De-Stain 15-30 minutes
2. Remove acetic acid with 5 x 20 minute washes with 50% EtOH
3. Incubate gel in Buffer #1 90 seconds, shaking vigorously by hand
4. Wash gel with dH2O 3 x 20 seconds to prevent background
5. Incubate gel in silver stain (Buffer #2) 1 minute shaking vigorously
6. Wash gel with dH2O 2 x 15 seconds
7. Develop with Buffer #3, watch for bands approximately 30 seconds 1 min
8. Stop with 10% Acetic Acid for 30 min
9. Wash gel with dH2O, gel is stable for a few weeks if you add 0.5% Acetic Acid



Solutions

50% EtOH
10% Acetic Acid

De-Stain Fix
50% MeOH
10% Acetic Acid
0.5 mL/L 37% Formaldehyde

Buffer #1 make once per month
0.13 g/L Na2S2O3 anhydrous
with dH2O

Buffer #2 make fresh each time
0.1g/50 ml AgNO3
1 ml/L 37% Formaldehyde

Buffer #3 once per month
60g/L NA2CO3 (note this NOT SODIUM BICARBONATE)
1 ml/L 37% Formaldehyde
30 ml/L in Buffer #1


courtesy of C.C. King - UCSD

Rb

Better to just buy a kit. I used to make my own reagents for years, but the commercial kits are faster, more sensitive, less variable and even cheaper when you figure in time.

The kits from Pierce and Invitrogen are the best.