``||´´_Linear_``||´ ´_Conformational_``||´´_Epitope_ ` `||´´_Retrieval

P.C. PC at no.email.sorry
Tue Feb 10 09:48:37 EST 2004


you mentioned that your deletions are tagged - the best control here is 
to blot with anti-tag Ab the same set of deletions. No one can ask for 
more.

It is likely not the case, but theoretically your epitope still can be 
conformational - protein can refold on the membrane. In fact, change 
from SDS (or more commonly sarcosyl) into Triton is one of the 
renaturation methods.

Does your antibody IP native protein? IP with your Ab, detect on the 
western with anti-tag and you shall be free of your intrinsic feeling!


Peter

redeamer wrote:
>>in a case like yours, when Ab works so well, it is definitely a linear 
>>epitope. My opinion - do not waste more of your time.
>>(you absolutely sure that you load at least as much or more of the 
>>deletion mutants on your gel for WB detection?).
>>
>>Peter
> 
> 
> Hi Peter,
> 
> almost sure... i could do a rough estimation from westerns with
> control proteins with the same length expo in HRP assay... but i do
> not quite believe that.. i do not like western in the sense of lacking
> quantitative characteristics, however qualitative ones are SUPERB:)
> 
> yesterday evening read one book on immunology, there was something
> like: "...somekind of interaction of native protein with somekind of
> linear epitope is crucial for immunity against native proteins..." i
> havent understood anything from that:) DOES ANYBODY KNOW how to
> decipher that, or i was a bit wrong in my partial citation, partial
> monologue...
> 
> intrinsic feeling is still inside me:(
> 
> ------------> Drew




More information about the Methods mailing list