P2D - silver stain

Michael Johns (johnsm@EM.AGR.CA)
Mon, 22 Mar 1999 16:31:54 -0500

After silver staining 2D gels, I get areas (about 1-2 sq cm) usually near the top, that are blank were there should be protein spots. I ran the same sample on a 1D gel and I get the same thing. Often the center of the lane is clear and the edges are quite dark. I realize that I should expect some background protein staining between bands, however, the background is very uneven. I don't to see this in coomassie blue stained gels. It seems like something is unevenly distributing itself in the lane and preventing the reduction of silver?

I have fractionated the samples to get 'pure' protein and have tried different sample buffers with no effect on the gel. Has anyone seen this in their gels? - are there any remedies or is this just a property of silver staining?

Michael Johns
Lacombe Research Centre
Agriculture and Agri-Food Canada
6000 C & E Trail
Lacombe, Alberta
Canada
T4L 1W1

Phone: 403-782-8100 ext. 268
Fax: 403-782-6120
johnsm@em.agr.ca
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