Best Chromatin IP antibody?(tag) - (May/08/2007 )

Hi ,
I will clone my gene of interest (to be IPed) in a vector with a suitable tag for ChiP. Then I will overexpress this plasmid in my cell line and then with an anitbody aproppriate for this tag I will IP it.
From your experience, what kind of antibodies work best in ChIP? (myc,HA, flag,V5,.....??)
Thank you for your suggestions.

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If you want to get the best signal to background ratio you should go for biotinylation of your protein so you can do stringent washes. Tagging your protein with biotin ligase recognition peptide and coepressing with biotin ligase will result in the biotinylation of your protein. You can get a very good signal to background ratio this way.

In our lab we have used GFP, HA, and FLAG tags all with similar signal to background as using antibodies for the specific protein of interest. None of these realy stand out above the others as far as the results go. It's just a matter of finding a tag that doesn't screw up the function of your protein.

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in my hands, the flag tag is the best...

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Hi!
Thank you for your comments. Could you please tell me which company you prefer for Flag antibody??

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