How to separate vegf dimer into monomer?

11 1
Mon Mar 3 12:39:43 EST 1997


In article <ziping-2802970933350001 at orphan.radonc.washington.edu>,
ziping at u.washington.edu (Ziping Yang) wrote:

> Hi, everybody
> I run vegf western and use B-mercaptoethanol (freshly added to lysis
> buiffer at 280-400 mM) as a reducing reagent. Samples were boiled before
> loading to gel. However,  VEGF dimer is always there and showed much
> strong band than monomer did. Any suggestions are wellcomed.
> ziping

We have had problems with VEGF dimers and multimers on SDS-PAGE as well. 
The problem seems to be worse with purified protein than with crude
extracts.  We now add 10% (v/v) beta-mercaptoethanol immediately before
boiling samples and include 1 mM DTT in the resolving gel.  This yields
only monomer on our Westerns.  Good luck.

-- 
Fern E. Murdoch
Biochemistry
Uniformed Services University
Bethesda, MD



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