I want to find binding sites of my transcription factor via ChIP-seq. I have a polyclonal antibody that weakly recognizes my protein in western (besides numerous other bands) and I am not sure if I can use that antibody for ChIP. How could I test that? I don't have any target candidates that I could use.
I am also thinking of tagging my protein in order to do the ChIP with a ChIP grade antibody against the Tag:
1. What is your experience with that?
2. Does overexpression of the tagged protein have negative influence on the ChIP assay? Should I use a weak promoter for that? Also the usage of a Tet inducible system would be possible; I would be able to titrate Dox for low expression levels.
3. Is a knockdown of the endogenous transcription factor neccessary?
Thanks a lot for you help!!!
Submit your paper to J Biol Methods today!
ChIP antibody question
No replies to this topic