making antibodies to low antigenicity proteins

Paul Bucciaglia paul-b at molbio.cbs.umn.edu
Mon Mar 7 16:14:03 EST 1994


In article <ming-050394103823 at sweetprotein.ahabs.wisc.edu> ming at ahabs.wisc.edu (Ding Ming) writes:
>In article <2l9mko$76l at mserv1.dl.ac.uk>, cooke at univ-perp.fr (Richard Cooke)
>wrote:
>
>> Bonjour,
>> 
>>         A colleague of mine is looking for references or methods for
>> raising antibodies against small, hydrophilic proteins. She has heard of a
>> method involving binding of IgGs to the protein. Any information would be
>> greatly appreciated.
>If you have IgGs for this protein, why do you bother to make Ab again? :)
>I think you were talking about coupling rather than binding, right?
>
>Anyway, for those small,  hydrophilic proteins, you probably need to
>conjugate them to some generic proteins like BSA, KLH or thyroglobulin.
>Simply treat them as synthetic peptide haptens. Another trick is to
>immunize animals with either SDS-PAGE or nitrocellulose powders if your
>protocol allows you do that.
>
>Ding Ming
>University of Wisconsin-Madison
>Telephone: (608)265-3544     Fax: (608)262-7420
>++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++

Beware of injecting nitrocellulose--it is antigenic!! I found out the hard
way.  If you do inject proteins bound to nitrocellulose (i first dissoved 
it in DMSO)  you can do your blotting to PVDF membranes but i've found PVDF
causes higher backgrounds than nitrocellulose when using luminescent detection.
Just my $.02.

paul b





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