ChIP from low amount of cells - (Sep/11/2006 )

Hi,

I would like to perform ChIP on chromatin from 5 x 10^5 cells.

Does anybody have experience with this? Should I use less antibody when I ChIP a low amount of chromatin (a few 100 ng perhaps)?

Thanks

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That amount is about 1/4 to 1/2 what people usually use so it's likely you could get away with scaling down the assay. Regarding your question about the amount of antibody to use, I'm guessing you suggest using less to lower the background. In addition to lowering the amount of antibody you should also decrease the amount of protein A beads since, in our experience, the beads contribute a lot to background.

You might also want to check out a method developed in Bryan Turner's lab where they perform ChIP on VERY low numbers of cells (Nature Genetics (38), 835-841. 2006).

Good luck,
Joel

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