P.C. PC at no.email.sorry
Sun Feb 8 20:26:20 EST 2004

how specific is your Ab in Western blot: does it detect your protein in 
a complex mixture, like cellular lisate?

is your purpose to locate the epitope or to produce an Ab for a 
particlular purpose?

A similar problem: people sometimes rely on far-western blot results to 
find interactors or locate regions of proteins involved in interaction. 
But there will always be, as you said, "some kind of intrinsic feeling". 
Anyone who has had an experience with a poor antibody or used a good one 
at higher concentration than necessary would agree. Anyway,there can be 
very many arguments for it and against it. With proper controls it is 

if your protein is fused with any sort of tag, you could always do a 
clean ELISA or a pull-down. If you are making deletions, some of the 
mutants might not be well-folded or even insoluble, so it is not always 
easy, of course.

good luck,


redeamer wrote:
> Hi guys!
> i have one, may be stupid, question: 
> If an antibody recognizes linear epitope, is there any sense to check
> this antibody on native epitope target? I mean if i do an epitope
> mapping and have a signal on my western blot (that i did from SDS
> PAG), should i do immunoprecipitation and then western to retrieve a
> possible conformational epitope for the same protein?
> I think that the answer is yes, but this is somekind of intrinsic
> feeling, that i cannot evolve now:(
> so i need YOUR help :)
> THANK YOU in advance,
> --------> Drew

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