Silver Stain Procedure for Slab Gels

Stock Solutions:

1. 0.36% (w/v) NaOH (0.36g in 100ml dH₂O)

2. 1% (w/v) citric acid

3. formaldehyde (37% w/v)

Working Reagents:

4. 0.8g AgNO₃ dissolved in 4ml dH₂O (in gradifrac tube)

5. 21ml 0.36% NaOH plus 1.4ml conc. NH₄OH (28%)

6. Stain Solution

add solution 4 dropwise into solution 5 while stirring,

then dilute to 100ml w/ dH₂O

7. Developer

2.5ml citric acid

0.25ml formaldehyde

dilute to 500ml w/ dH₂O (half recipe for one gel)

Procedure

1. Soak gel in 50% MeOH for at least 2.5hrs. (OK O/N)

2. Rinse gel and container w/ dH₂O, then add stain. Incubate 15 min w/ shaking

3. Rinse gel 5 min w/ several washes of dH₂O

4. Add developer and incubate 10 min w/ vigorous shaking or until band develops

5. Pour off developer and rinse w/ dH₂O. Stop with 50% MeOH. (MeOH stops the staining process slowly. Perform this step just prior to anticipated endpoint.)