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        The first thing I want to know is what type of protein your antibody
was raised against.  Is it native or denatured or is it one of the
bacterially produced peptide sequence?  Also are you working in the same or
cross species?  A majority of antibodies raise against denatured protein
species will not react against native proteins as fixation does not
necessarily denature the protein, particularly if it is a monoclonal and not
polyclonal.  Yes it will work in a western blot where you are working with
denatured proteins.  The question is whether it will immuno-percipitate,
where you have native protein.  The binding site of the antibody may have
been uncovered during its denaturing process.  You would then have to treat
your material in the same manner.  A while back I had a student working with
a protein that was denatured with 6m urea and his antibody raised against
that.  He could not get any immunolabeling until I told him the treat his
cells with urea.  The cells then lit up like a Christmas tree.

Joe Goodhouse
Confocal / EM Core Laboratory
Department of Molecular Biology
Princeton University
609-258-5432

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