Protein Electrophoresis

marc abrams abramsm at erols.com
Fri Sep 4 07:57:53 EST 1998


To all contributors:

    Thanks for all your help! Your suggestions are appriciated.
I have tried running l o n g e r  gels (up to 36cm) with very little, if
any, improvement in the separation.
    I am going to try a gradient gel containing 4-8M urea.  I'll let you
know what happens.  I may try the tricine gel again as well.  I think a
native gel and IEF are reasonable ideas as well. If none of it works,
at least I'm learning fun new techniqes :)
-marc
marc abrams wrote in message <6skk83$si6$1 at winter.news.erols.com>...
>    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
>
>





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