Immunoprecipitation protocol

Alexander P Rostovtsev rosto001 at maroon.tc.umn.edu
Fri Jul 22 15:39:39 EST 1994


In <bcguilf.25.0 at muccmail.missouri.edu> bcguilf at muccmail.missouri.edu (Guilfoyle Lab) writes:

>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.

	Why do you use protein A-Sepharose binding monoclonal Abs rather
poor? I usually use monoclonal Abs crosslinked to the protein G-Sepharose
with dimethyl pimelimidate - very reliable and stable immunosorbent.
Another advantage is almost 100% regeneration so you can reuse the same
sorbent several times.
Alexander Rostovtsev 



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