Alternatives to Laemli?

MICHAEL L SULLIVAN mlsulliv at facstaff.wisc.edu
Mon Feb 16 11:53:04 EST 2004


Several years ago, I wanted to run a gel at lower pH to maintain an ester linkage between 2 proteins (see Sullivan and Vierstra, J. Biol. Chem. 268:8777)  A colleague suggested the method in the following reference:

Weber, K, and Osborn, M (1969) The reliability of molecular weight determinations by dodecyl sulfate polyacrylamide gel electrophoresis.  J. Biol. Chem. 244, 4406-4412.

The gel gets run at pH 7.2 in a phosphate buffer.  Since the original reference is old, it used tube gels, but we adapted it to run slab gels.  Since there is no stack with this one, sample volumes need to be small to get tight bands.

I hope this helps.

Mike Sullivan
Research Molecular Geneticist
USDFRC-ARS-USDA


----- Original Message -----
From: ahernan at cica.es
Date: Monday, February 16, 2004 10:25 am
Subject: Alternatives to Laemli?

> Hi all,
> 
> I am trying to run a protein-RNA complex on a SDS-PAGE gel but I 
> was 
> told that laemli's high pH can promote RNA hydrolysis. Any one 
> knows 
> of an alternative buffer system for protein SDS-PAGE at (near) 
> neutral pH? I know Novex sells them as precast (and so do BioRad 
> and 
> Amersham...so it must not be under a patent) but buying such 
> expensive stuff at this stage is out of the question.
> Thanks a lot.
> Cheers,
> Agustin
> ---
> 

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