How to count labeled protein in SDS-PAGE gel
John E. Wiktorowicz, Ph.D.
johnw at lynxgen.com
Mon Oct 23 15:05:19 EST 2000
Yes, peroxide at high temp works. Another approach is to incubate the gels
in ammonium hydroxide at high (~60°C) to hydrolyze the protein and extract
the radioactivity from the slices. Use the same trick and digest in
scintillation vials. The ammonium can be evaporated and scintillation fluid
added after digestion. Forgot to ask, but this assumes you're trying to
count tritium and C14. Iodine125 of course can be counted directly (gamma
emmiter), and S35 may be high enough energy to count without extraction.
> From: "lxh" <lxh at dartmouth.edu>
> Organization: Dartmouth College, Hanover, NH, USA
> Newsgroups: bionet.molbio.proteins,bionet.molbio.methds-reagnts
> Date: Sat, 21 Oct 2000 18:16:01 -0400
> Subject: How to count labeled protein in SDS-PAGE gel
> Does any know a way to count the radioactivity of proteins from a SDS Gel
> stripe? What is the best way to extract the protein or dissolve the gel? I
> assume it is not a good idea to count it directly in a scintilation counter.
> But the extraction protocol I 've seen takes too many steps, and I am afraid
> I will lost some proteins since I only have several microgram protein per
> band. Thanks.
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