Silver staining polyacrylamid gels -Reply Correction

Kathy Mangold KMANGOLD at MAIL.MCG.EDU
Fri Oct 17 10:45:27 EST 1997


(Added time in step 3)

I've used this method for years with great success:  Bassam et al.,
Analytical Biochemistry, 196:80-83, 1991.

Briefly:

1.  Fix gel in 10% acetic acid for 20 minutes
2.  Rehydrate gel with 3 x 2 minutes in dH2O
3.  Saturate with AGNO3 (1 g/liter with 1.5 ml 37% HCOH/liter) for 30
minutes
4.  Briefly rinse off the surface AgNO3 with dH2O
5.  Develop with FRESH developer (30 g/liter, 1.5 ml 37% HCOH/liter,
2 mg/liter Na2S2O3-5H2O); watch for bands to develop in 2 to 5
minutes
6.  Stop development with 10% acetic acid for 5 minutes
7.  Rehydrate gel in dH2O before drying

Silver nitrate solution can be reused for several weeks, although
intensity does diminish over time, but reusing means less to dispose
of properly.  I use fresh developer every time.  100 ml of silver
nitrate is plenty for minigel setup (10 cm x 8 cm), and ~40 ml per
for each development.  (Since I usually do 2 gels a day, I make up
100 ml of developer also.)

I hope this is helpful; let me know if more info is needed.

Kathy A. Mangold, Ph.D.
Anatomic Pathology
Medical College of Georgia
Augusta, GA 30912-3605





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