This is part of my undergrad project that involved studying a protein.
As part of my project, i want to study whether my protein will bind and acetylase core Histone or not. I have clone, purified the interested protein. I also had Histone H4,H3 in stock.
What I want to do is to design an experiment where my protein will bind to core histone by using co-IP approach. Here is my thought. Please comment.
Mix my protein with Histone in appropriate buffer
pull one protein with antibody, later with agarose bead
finally, detect with histone antibody
do a WB to confirm size change.
I did my control experiment first with no antibody binding first, but i got high background that will mask the result later. I would like everyone to comment on this traditional co-IP approach. Does it worth to buy the commercial kit or can I still improve the protocol somehow?
Help design a protein-protein interaction using co_IP
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