I got a problem with my I.P. I'm trying to immunoprecipitate a FLAG tagged protein from a transgenic worm lysate (total lysate) using anti-FLAG Ab-ProteinG-sepharose. When I analyze equivalent amounts of total lysate before IP, bead extract, total lysate after IP through western blotting, almost all (at least 90%) the tagged protein bands were disappeared in lysate after IP, while strong in lysate before IP. However I couldn't see ovbious band either in the bead extract. My question is 'Where is the disappeared bands?'
Submit your paper to J Biol Methods today!
Problem in immunoprecipitation
1 reply to this topic