Silver staining with ampholytes
Dress at biosci.arizona.edu
Tue Mar 15 11:46:30 EST 1994
In article <199403142310.PAA05895 at net.bio.net>, jproudman at ASRR.ARSUSDA.GOV
("JOHN PROUDMAN") wrote:
> Hi, Lyndon-
> I do silver staining of both SDS gels and IEF gels. The only basic difference
> between the two procedures is that with IEF, I first fix the proteins in
> 20% TCA. Then the gel is washed in 10% ETOH-5% HAc (which is my first step
> for an SDS gel). Pharmacia sells a silver stain kit which I have used for
> both of these procedures, but I now make up my own reagents and they work
> just as well. I suspect that your problem may be solved by first fixing
> your proteins with TCA, then using your current silver stain procedure.
> Hope this helps.
There's a nice protocol for silver-staining 2-D gels in the Hoefer catalog.
It's been a long time but I think the fixation is very similar to what
describes above, and the fixation is probably the important parameter here.
Pharmacia also describes a silver-staining technique in the Phast-gel
manual, they may have a separate technical bulletin they'd send you
not for free, they are real tightwads). But the Hoefer catalog is free and
their technical services is great. Give them a call if you have problems
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