Silver Staining

Dominik Haudenschild, 10-28-94 at CBRC

-Clean all plastic-ware carefully. Wear Powder-free gloves. Use Milli-Q water whenever water is needed.

-Coomassie stain/destain gel if needed, or fix in 50% Ethanol/10% Acetic Acid

for 30 minutes minimum

1) Soak twice 20 minutes in water to remove Acetic Acid

2) Soak in 2% Glycerol, 40% Methanol, 10% Ethanol 20 minutes to

dehydrate and stabilize gel

3) Stain for 20 minuts in 100ml stain

NaOH 187. 5ul of 10N or 0.075g in 21ml water

NH4OH(30%) 1.4 ml

20% AgNO3 4 ml (or 0.8g in 4ml water)

*Add silver dropwise, while stirring, allowing solution to clear before

adding more silver.

4) Wash minimum of 3 times 2 minutes in water. Longer washes mean

less background.

5) Immediately before use make developer.

0.5% Citric Acid 1 ml

37% Formaldehyde 51.2 ul

Milli-Q to 100ml

Develop until ready. 4 minutes is average, longer than 15 minutes

means something is wrong. Can be re-done by washing gel

many times with water, then going back to step 2.

6) To stop reaction, just wash well in water. Dry gel as usual.