help with silver stain

XZHENG at GAMMA.IS.TCU.EDU XZHENG at GAMMA.IS.TCU.EDU
Tue Oct 24 15:47:52 EST 1995


Hi, there:
I really appreciate it if somebody can help me solve this problem. I tried to
use silver stain to visualize protein bands on acrylamide gels, but I got a
high background(black vertical streaks and dark brown background). the method
I used was developed by Merril and co-workers as follows:
1. fix the proteins in the gel in 40%methanol, 10% acetic acid for 30min
2. fix twice in 10% ethanol, 5% acetic acid for 15min
3. soak the gel for 3-10min in fresh oxider solution (0.0034M potassium
dichromate, 0.0032 N nitric acid)
4. wash the gel 3-4 times in water until the yellow color has been washed out
5.
  soak the gel in fresh silver reagent (0.012 M silver nitrate) for 15-30 min
6. wash the gel with water for 1-2 min
7. wash the gel for about 1 min in developer (0.28 sodium carbonate, 1.85%
paraformaldehyde)
8. replace the developer with fresh solution and incubate for 5 min
9. replace the developer a second time until satisfactory staining has been 
obtained.
10. stop development with 5% acetic acid
Any suggestions on how to reduce the background? thanks in advance!
Alan Zheng
Chemistry Dept.
Texas Christian Univ.



More information about the Methods mailing list