Myc-tag in ChIP - problems with ChIpping myc-tagged proteins (Aug/22/2005 )
I'v been trying for a while to ChIp myc-tagged transcription factors. I don't get recoveries significant higher than background, whereas we know that at least some of them are present on the promoters we PCR (because we get high recoveries for the corresponding endogenous proteins with antibodies directed against epitopes in the proteins themselves).
I want to use the myc-tag to make faire comparisons, so that i don't have the problem of using different antibodies.
Has anyone ever used the myc-tag successfully in ChIP, or does anyone have an explanation / suggestions? Thank you very much!
You can increase washing rounds or using an alternative washing buffer, which is more stringent.
Hope that will increase the difference between your control and ChIP DNA.