[Q] 18% SDS-PAGE

Rich Dudley rdudley+ at pitt.edu
Fri Jun 18 12:13:20 EST 1999


Namjin Chung wrote:

> I'm trying to work on 18% tris-glycine SDS-PAGE for 7-9kDa proteins.
> Somehow, when I run the gel, dye smears broadly behind buffer front.
> Also, western blot shows smeared bands.  Why does this happens?
>
> Namjin Chung
> Department of Pharmacology & Cancer Biology
> Department of Genetics
> Howard Hughes Medical Institute
> Duke University Medical Center

Another thing to try is using Tris-tricine buffer and Amresco's
PAGE-Plus acrylamide.  I've used it for proteins that small at the 14%
or 16% range.  Just make the gel using the standard recipe (adjusted for
% of the stock sol'n), and substitute the PAGE-Plus for the acrylamide.

The traditional methods don't resolve proteins that small very well.  At
that range, the pore size of the polyacrylamide doesn't sieve well, and
your proteins are jumbled around with the SDS micelles.

rich

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