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Immunoprecipitation protocol

W. Lee Adair acannons at com1.med.usf.edu
Mon Jul 25 10:34:40 EST 1994

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.

Andy Cannons
 Guilfoyle Lab 
(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.

: Thanx,

: Tim Ulmasov
: Biochemistry,
: University of Missouri-Columbia
: bichguil at mizzou1.missouri.edu

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