I've never done IP and would appreciate any feedback.
I hypothesize that portein A interacts with protein B. All my cancer cell lines show low levels of A and all show high levels of B. Thus, I was planning to overexpres protein A in one of my cancer cell lines, pull it down it with the antibody and see if it interacts with protein B? However, now i'm having doubuts whether I've chosen the right cancer cell line. All my cancer cell lines show low levels of protein A but only a normal immortalised cell line shows high levels of A and B both? So, should I have used the normal immortalised cell line instead of my cancer cell line?
The other reason why i've used this cancer cell line is that overexpression of protein A showsn tumor suppressive properties by in vitro and in vivo assays and I thought it would be interesting to check the interaction in this cancer cell line instead?
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