BMB anti-HA and immunofluorescence

Tien-Hsien Chang tchang at magnus.acs.ohio-state.edu
Tue Feb 14 16:57:52 EST 1995


     After attempting a few channels to obtain anti-HA antibodies sold by
BabCo, we are still unable to get any.  Thus, we were forced to use mouse
anti-HA monoclonal from BMB.  This (expensive) antibody, although useful in
Western analysis on our hands, appeared to have significantly high background
in indirect immunofluorescence studies.  In several tries, even using
preadsorbed (by yeast powder) BMB anti-HA, we still saw pretty obvious
background staining in the negative control, i.e. strains which do not contain
any HA-tagged proteins.  Our control experiments using different antibodies
lighted up where they supposed to be.  So, the problem seems to be derived from
BMB's anti-HA per se.

     Is there anyone out there successfully using BMB's stuff for indirect
immunofluorescence studies on S. cerevisiae?  If so, can you kindly offer
suggestion to us?  Also, can someone who has BabCo's anti-HA kindly offer a bit
of that antibody?  Thanks in advance.

                              Tien-Hsien Chang
                              Department of Molecular Genetics
                              The Ohio State University
                              484 West 12th Ave.
                              Columbus, OH 43210

                              (614)-292-0631
                              (614)-292-4466 [fax]
                              chang.108 at osu.edu



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