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western blots

Unknown Unknown
Fri Nov 3 07:18:51 EST 1995

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

I suppose that specific binding  implies a high affinity binding while
non-specific binding is of low affinity. Therefore the high affinity
binding will decrease much more rapidly than the non-specific binding
when you add excess peptide while staining the western blot.

In my opinion adding peptide to the primary antibody solution is
a good control because when your band vanishes you can be sure that
that antigen  has the same epitope as the peptide.


dirk.jochmans at chaos.kulnet.kuleuven.ac.be

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