You could try treating your mixes with PAS before adding your specific
antisera, pellet the beads & then add antisera to the sup, then add PAS
again. This should pre-absorb out all the non specific binding. You
could also try pre-immune serum as a pre-incubation.
(bcguilf at muccmail.missouri.edu) wrote: : Hi,
: I'm trying to study some protein-protein interactions using in vitro
: translated HA-epitope-tagged proteins. I need a reliable protocol for
: doing immunoprecipitations from these in vitro translation reactions
: using monoclonal 12CA5 which recognizes HA epitope. So far it looks like
: I'm having high background problem coming from non-specific binding to
: Protein A-Sepharose beads.
: Any suggestions, references or hints would be greatly appreciated.
: Tim Ulmasov
: University of Missouri-Columbia
:bichguil at mizzou1.missouri.edu