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!
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!!!!!
--- --- --- -- -- -- --- --- ---
Richard J. Dudley (rdudley+ at pitt.edu)
Research Specialist V
Dept. of Cell Biology and Physiology
University of Pittsburgh
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