Silver staining polyacrylamid gel

Administrador de iihema.fmed.uba.ar postmast at iihema.fmed.uba.ar
Mon Oct 20 18:44:47 EST 1997


You could add a step between 2 and 3 that's supposed to diminish background:
1 minute treatment with 0.02% sodium thiosulfite.  Must have the ref. 
somewhere, only I just can't find it (I'll dig deeper only if necessary).
Also, I've reused silver nitrate solution for almost a year and it works.

Facundo Garcia Bournissen.
postmast at iihema.fmed.uba.ar

>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 Na2CO3, 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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