Koshland Web Site/Methods
Silver Stain of Protein Gels
  1. Fix gel in 50% methanol for greater than 1 hour.
    rinse gel in dH2O 3 times 5 minutes each
    place back in 50% methanol for 1 hour.
  2. Stain gel in 100 mls of stain solution for 30 minutes
  3. Rinse gel in dH2O 3 times 5 minutes each.
  4. Place gel in Developer until bands are almost as dark as desired,
    usually 20' - 2 hours.
  5. Stop developing and fix stain by placing gel in 50% methanol. Bands may continue to darken in methanol overnight, it's best to photograph gel immediately.
  6. Photograph on light box. (for type 57 film use settings of 1/125 sec, f=22; for type 55 film use settings of 1/15 sec, f=11)
  7. Wrap gel in prewet BioWrap® and allow to dry overnight

Stain solution
dissolve 0.8 g of AgNO3 in 4 mls dH2O
add a mixture of the following....
21 mls of 0.36% NaOH plus1.4 mls of concentrated NH4OH (a little extra may be needed; add just enough to get silver to dissolve wait for lag!) then bring up to 100 mls with dH2O (by adding 72.8 mls)

500 mls of dH2O
2.5 mls of 1% Citric Acid (monohydrate!)
0.25 ml of 36% formaldehyde

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