How to separate vegf dimer into monomer?
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.
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
Uniformed Services University
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