MAbs and Western Blot.

Ian A. York iayork at panix.com
Mon Jan 22 09:50:57 EST 2001


In article <ZRWa6.2730$1K1.24674 at newsr1.maine.rr.com>,
Joe Chandler <jchandler at mainebiotechnology.com> wrote:
>In the course of our development work, we have generated scores of MAbs.
>They work in beautifully in EIA and IHC.  However, we seem to find that even
>high binding MAbs do not often work well in Westerns.  I have done a

That's quite common, depending on the antigen.  In western blots, the
antigen is at least partially denatured (sometimes it partially renatures
after binding to the nitrocellulose, but rarely do you get complete
folding once again).  If your mAb tend to be conformation-dependent (if,
for example, you're looking at antigens that are disulphide-bonded, or
mulitmeric) then they'll usually work less well on Westerns.

IHC also can present conformation problems because of fixing, but these
are generally of a different kind, and generally less severe.

Ian 
-- 
    Ian York   (iayork at panix.com)  <http://www.panix.com/~iayork/>
    "-but as he was a York, I am rather inclined to suppose him a
     very respectable Man." -Jane Austen, The History of England






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