Silver Staining of Proteins in Gel Author: Sharad Purohit
Source: Contributed by Sharad Purohit
Date Added: 2002-05-14
Date Modified: 2004-04-29
Fixative Solution: 50% methanol, 10% Acetic acid, 0.25% Glycerol
Solution A: 2% silver nitrate and 2% ammonium nitrate.
Solution B: 10% tungstosilic acid
Solution C: 3% formaldehyde
Developer solution: 5% sodium carbonate
Stopping reagent: 5% acetic acid.
- Fix the Gel in the fixative solution for 20 minutes
- Wash the gel in deionised water 2 times for 10 minutes each.
- Mix 1 ml each of solutions A, B, and C and 7 ml of deionised water and just before use add 10 ml of developer solution stain the gel in above solution in a zipper bag till desired contrast is obtained. it usually takes 10- 15 minutes depending on the room temperature
- When desired contrast is obtained, drain the staining solution and quickly add the stop solution to stop the staining.
- Handle the gels with gloved hands only.
- Use deionised water.
- Treat the glassware with concentrated nitric acid before use, it is preferable to use polyethylene zipper bags for the full procedure.
- This receipe is for one 0.75mm thick gel, increase the quantities as per the thickness of gel.
- To destain the gel put it in 3% hydrogen peroxide
This protocol is based on the method of Gotlieb and Chavko
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