Protein Electrophoresis

Peter pxpst2 at unixs.cis.pitt.edu
Thu Sep 3 10:12:03 EST 1998


In article <6skk83$si6$1 at winter.news.erols.com>, "marc abrams"
<abramsm at erols.com> wrote:

>     I have been using a high-percentage acrylamide SDS-PAGE system to
> resolve a 25kd protein and its modified form.  A post-translational
> modification causes a conformational change which creates a "shifted"
> species of the protein in Western blot experiments.  I would like to try
> different gel systems to see if I can better resolve these two species of
> the protein (increase the migration distance difference) so that the result
> is unambiguous.
> Does anyone know of gel systems that have been used to
> separate subtle difference in a protein based on conformation?
> I tried Tris-Tricine and Bis-Tris gels as well, with no success.

What do you mean by conformation?

I run continuos Tris tricine gels(Schlaeger et al) and can easily seperate
a 32 kd glycosilation variant of my protein from a 34 kd form.  I have
also been successful at seperating much smaller fragments(~2 kd from 5
kd).

Peter

-- 
"Don't you eat that yellow snow
            watch out where the Huskies go"    FZ

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