On Fri, 16 Feb 2007, . wrote:
> "atahualpa" <botulina from poczta.onet.pl> wrote in message
> news:Xns97C8BB8867C05sralwaspiesgazetapl from 193.42.231.152...>> Hi!
>> I have a following problem with immunoprecipitation:
>> When I develope western of the cell extract with the antibodies for the
>> protein I am dealing with I see the protein that I expect and two, much
>> thicker, bands higher. I want to immunoprecipitate them and go for MALDI.
>> I
>> have run immunoprecipitation of the cell extract with the same antibodies
>> and I run gel for coomasie staining. I can see on it very nice band of my
>> protein and nothing more beside background. It seems that IP works since I
>> can catch my protein, but where are the other two guys? Why they are not
>> caught in IP although very nicely detected on western with same
>> antibodies?
>> The only reason that comes to my mind is that western is developed in
>> conditions different from IP (milk vs lysis buffer). Could it be that
>> lysis
>> buffer (contains 1% NP40) affects binding conditions of those two not
>> affecting binding of the "original" protein?
>> I would appreciate all suggestions.
If I understand you correctly, could it be that your antibody will detect
the reduced forms in western blot, but not the native forms?
