Tris-Acetate SDS-PAGE gels

Peter pxpst2 at SPAM.SUXS.unixs.cis.pitt.edu
Sat Jul 25 11:52:18 EST 1998


In article <6parrc$cqt at gazette.bcm.tmc.edu>, fjetland at bcm.tmc.edu (Conrad
R Fjetland) wrote:

> Hello,
> 
>         I am trying to separate two proteins on an SDS-PAGE gel of very
> similar molecular weights.  The first is glutathione reductase with a MW
> of 55 kD while the second is a mutant GR protein that is 57 kD.  We would
> like to monitor their expression in CHO cells using an anti-GR antibody.
> I have found a premade gel by Novex that is a tris-acetate gel which has
> the separation range that we are looking for.  However, their gels will
> not work with our BioRad gel box.  Has anybody worked with tris-acetate
> SDS-PAGE gels before?  If you have, could you please give me the
> protocols.  Thank you very much in advance.

You might want to try to use a Tris-Tricine gel.  It is best for
seperating proteins that differ by as little as 1 KB.  I would suggest
starting with a 10 percent gel and wok up to a 13  % gel in 1/2 percent 
increments.  This system will increase the resolution of proteins that are
less than 100 KD w/o requireing a gradient gel.
the refernce for this procedure is:
H. Schagger and G. Von Jagow (1987) Anal.  Biochem. 166, 368-379


Good luck and I hope this helps

Peter

-- 
"Don't you eat that yellow snow
            watch out where the Huskies go"    FZ

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