Problem with Native PAGE

Ding Ming ming at ahabs.wisc.edu
Tue Oct 4 12:45:53 EST 1994


In article <Cx4wwp.Dy3 at serval.net.wsu.edu>, pxu at wsuaix.csc.wsu.edu (Pin
Xu) wrote:

> I recently tried to purify an enzyme on native PAGE. I used Bio-Rad mini-gel
> apparatus and Tris-Glycine (Laemmli) buffer system (without SDS). The problem
> is I can never get a clean gel: vertical streak, band diffusion, protein
> precipitate in the interface bewteen stacking gel and separating gel, etc, etc
> etc. I wonder if there is a good protocol for Native PAGE?

I wonder if your enzyme has a basic pI which make the protein precipitate
at pH 8.8. You need to clarify this by a titration experiment. If you do
deal with a basic protein, you have to used an acid system, e.g.,
acetate-KOH or others which can be found in almost any PAGE book.

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Ding Ming
University of Wisconsin-Madison
Telephone: (608)265-3544     Fax: (608)262-7420
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