immune precipitation

FGARBRECHT at mis.mcw.edu FGARBRECHT at mis.mcw.edu
Sat Oct 3 21:55:43 EST 1992


If you stain your gel for protein before autoradiography,
and you see bands in the appropriate places for immunoglobulin,
that suggests that the monoclonal is indeed binding the SPA and is
being precipitated, and that you should look elsewhere for the
problem. One difficulty may be that the protein you are trying to
immunoprecipitate does not label efficiently under the conditions you
are employing.  For 35S-methionine labeling, make sure the protein
contains methionine!  You may also extend the labeling time overnight
to enhance labeling (at the expense of increased background).  Check
out "Current Protocols in Immunology" for detailed protocols and
troubleshooting tips.  Good luck.

Fred Garbrecht
Medical College of Wisconsin



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