SILVER STAINING
FOR PROTEIN IDENTIFICATION BY MASS SPECTROMETRY

  1. Fix gel in 40% methanol/10% acetic acid for several 10-15 minutes changes or overnight.   Use a clean container and cover to exclude airborne contaminants.

  2. Rinse gel in deionized water.

  3. Sensitize with 0.02% sodium thiosulfate for a total of 2 minutes.
    Note: make solution fresh that day

  4. Rinse gel and plate holding the gel 2x with deionized water (1 min per rinse)

  5. Incubate with 0.1% silver nitrate at room temperature for 30 minutes.

  6. Rinse gel and plate holding the gel 2x with deionized water (1 min per rinse)

  7. Develop gel in 0.04% formalin/2% sodium carbonate (~4-8 minutes)

  8. Stop development with 1% acetic acid.   Do this quickly, rinse about 1-2 minutes.

  9. Apply fresh 1% acetic acid for gel storage.