Berglund:Silverstaining

Adapted from Shevchenko et al, 1996,  Anal. Chem. 68: 850-858 (Adapted by Protein Research Group- Dept of Mol. Bio. -Odense University- Denmark)

Materials
destaining solution- Methanol:acetic acid: water (45:5:45) sensitizing agent 0.02% sodium thiosulfate- 0.1g Na2S2O3 per 500ml water 0.1% silver solution- 0.1g in 100ml water Developing solution- 150µl 37% formalin solution, 12.5g Na2Co3, 500ml water Quench solution- 5ml acetic acid in 500ml water

Methods

 * 1) Fix gel with destaining solution for 20-30 minutes
 * 2) Rinse gel with water for 20-60 min to remove acid
 * 3) Sensitize gel with 0.02% sodium thiosulfate for 1-2 min
 * 4) discard solution and rinse gel with two changes of water (1 minute each)
 * 5) incubate gel in chilled 0.1% silver solution for 20-40 minutes at RT
 * 6) discard solution and rinse gel with two changes of water (1 minute each)
 * 7) Develop gel with Developing solution with shaking (replace developing solution when it turns yellow)
 * 8) Quench when sufficient staining is obtained by discarding developing solution and adding quench solution
 * 9) store in 1% acetic acid at 4°C or dry gel