western blots

[Stephen R. Lasky]

In article <47382l$opc at netserv.unmc.edu>, dmlepley at netserv.unmc.edu
(Denise Lepley) wrote:

> Hello all!  I was wondering what the consensus is for trying to use the 
> peptide antigen to compete off non-specific primary antibody binding for 
> western blot analysis.  In my opinion, using the same peptide the 
> antibody was raised against will compete off non-specific as well as 
> specific antibody binding.  So, in effect, adding peptide to the primary 
> antibody solution is a meaningless control.  What are your opinions on 
> this?  

Theoretically, use of a specific peptide should not compete non-specific
binding.  From my experience using this technique to identify which bands
are actually Jun or Fos proteins, it works.  

SRLasky

-- 
Stephen R. Lasky Ph.D.
Section of Experimental Therapeutics
Roger Williams Medical Center
Providence, RI 02908
Phone: 401-456-5672      Fax: 401-456-6569
e:mail: Stephen_Lasky at brown.edu