Protein Electrophoresis

Rich Dudley rdudley+ at pitt.edu
Thu Sep 3 07:43:17 EST 1998


Conformation?  Proteins don't usually separate according to conformation.
Glycosylations, etc. add actual mass, and phosphorylations adjust the charge
balance (thereby adjusting the apparent MW).

What is the modification?  I would think a good tris-tricine would work, but
that might take some tinkering.  A native PAGE might also work.

Hope this helps!

rich

marc abrams 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.
> Thanks in advance!!!!!
> -Marc



--
--- --- --- -- -- -- --- --- ---
Richard J. Dudley (rdudley+ at pitt.edu)
Research Specialist V
Dept. of Cell Biology and Physiology
University of Pittsburgh
http://www.cbp.pitt.edu
---> search BIONET archives at http://www.bio.net <---

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