IUBio

gel electrophoresis of peptides...

Krieger Lab dresnick at mit.edu
Thu Jul 11 05:11:56 EST 1996


> In article <4s19qb$guv at deft.cc.purdue.edu>,
>    wilkinst at deft.cc.purdue.edu (Thomas Wilkinson) wrote:
> -->Hello,
> -->
> -->I was wondering if somebody had some experience with SDS gel electrophoresis of
> -->peptides.  I have a sample that I would like to analyze to see if it contains
> -->a short (12-residue) peptide, molecular weight = 1462.
> -->
> -->The sample is not some mixture of many species...the sample to be analyzed
> -->either has the 12-mer, or else the sample contains no peptide whatsoever. Thus,
> -->this gel should be really easy to interpret.  I also have a similar sample that
> -->I would like to analyze for peptide content, except in this case I am looking
> -->for the presence or absence of a 33-residue peptide (MW = 3487).
> -->
> -->I would like to run these samples out on an SDS gel, and then coomassie-stain.
> -->My question is that I don't know what the optimal percentage of polyacrylamide
> -->would be for this task.  What percent gel is best?  Is gel electrophoresis of
> -->peptides commonly done?  Your suggestions are much appreciated.
> -->
> 

Sounds like your peptides are too short to do SDS-PAGE with.  I'd recommend either:

1) Reverse phase HPLC (assuming you know where they run), or

2) Mass Spectroscopy

I'd think that either of these would be preferable to PAGE....


-David




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