Hi! Has anyone tried to ChIP either histones or sequence-specific transcription factors without prior crosslinking? How do the results compare?
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Posted 09 August 2010 - 09:58 AM
Serendipitous, on 08 August 2010 - 04:52 PM, said:
Hi! Has anyone tried to ChIP either histones or sequence-specific transcription factors without prior crosslinking? How do the results compare?
You should look up Native ChIP. As for experiences, a member of my old lab did native ChIP for some histone mods (don't remember which anymore) and got good results (not too much different from using crosslinked chromatin). This works because histones are so tightly bound to the DNA. TFs and other DNA bound proteins may not be bound tight enough to hang on through detergents and sonication and I don't know, personally, of anyone who has tried native ChIP for these types of proteins.
