Co-immunoprecipitation of nuclear proteins

ege ege at mit.edu
Thu Feb 6 16:42:25 EST 1997


Uhh, RIPA, NP40/DOC are likely going to disrupt many protein/protein
interactions.  Try something milder like ELB (E1A lysis buffer described
many years ago in a paper with Ed Harlow as an author looking at E1A
associated proteins)  If you see the bait and prey coming down
individually you don't have an extraction problem.  If you don't see them
coming down together you're likely wasting your time on an in vitro
artifact of the 2 hybrid.  Maybe your just really unlucky and your
antibodies recognize epitopes masked by the bait/prey interaction.



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