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Hey All,
I have been trying to co-IP two transcription factors from nuclear fractions. Up until today, I have had varying success with the 'trap' protein, but almost NEVER see my original "bait" protein that I IP'd for. I can clearly see it in the same samples that have not been IP'd, and since I can see heavy chain and some interacting partners, it appears to have IP'd, but I can't get it to show up on an immunoblot. Problem number 2 is that now my trap protein appears to me migrating 20 kDA higher in the IP samples than in the untreated lysates. Any ideas on either front?? Thanks!
