resolution on PAGE??

Chris LaRosa clarosa at biocomp.unl.edu
Wed Mar 22 09:43:23 EST 2000



Ben Gantner wrote:
> 
> Hi all,
> 
> Does anyone have protocols or good web sites that discuss methods for
> improving resolution of SDS-PAGE?  I'm trying to get separation of a
> fairly complex mixture, and I'm not getting particularly good
> resolution.
> 
>
Without more detail on your particular application its hard to help you
.
Having run hundreds of SDS-Page on plant proteins some years ago, here
are a few words of advise:

Use a good cooled set up like biorad protean II

Run gradient gels.

I suspect that the precast gels they sell now days may be a good
investment, but never had those.
Use fresh acrylamide, from good source, filtered through .44 um filter

Sample preparation is everything.   Use proteinase inhibitors.  Work
fast , keep samples cold,  spin the samples hard to remove
particulates.




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