gel electrophoresis of peptides...

David Martin damartin at bioslave.uio.no
Thu Jul 11 04:07:52 EST 1996


In article <4s19qb$guv at deft.cc.purdue.edu>,
   wilkinst at deft.cc.purdue.edu (Thomas Wilkinson) wrote:
-->Newsgroups: bionet.molecules.peptides
-->Subject: gel electrophoresis of peptides...
-->Summary: 
-->Followup-To: 
-->Distribution: world
-->Organization: Purdue University Computing Center
-->Keywords: 
-->
-->
-->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.
-->

You are at the lower limit for SDS-PAGE and would need to use a Tris-Tricine 
buffer system (schagger & von Jagow, Anal. biochem. 1987, 166, 368-379)
and a fairly high percentage gel (15%-20%) Even then the 12mer may be too 
small.

Alternatively, if you just wish for a yes or no answer for the presence of 
peptide, you could dot your sample onto PVDF and stain with amido black.

best of luck.

.d

* David Martin, PhD -  Post-Doctoral Research Fellow        *
* Atherosclerosis and Thrombosis research group             *
* Biotechnologisenteret i Oslo                              *
* Postboks 1125 Blindern, N-0316 Oslo, Norway               *
* Tel: +47 22 95 84 54  Fax: +47 22 69 41 30                *
* http://www.uio.no/~damartin/  david.martin at biotek.uio.no  *



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