To charaterize TF function in specific bone marrow or immune cell subsets, we only have relatively small cell numbers (106 or less) to prepare chromatin from. Does anybody have (good) experiences with high sensitivy ChIP kits or home-made protocols for small populations? Cheers and Thx
Hello
Protocol details in the attachment . i hope help you
Good Luck
http://www.biomedcentral.com/1471-2164/16/46
Thanks a lot for the helpful protocols, Saleh! We are not sure yet if it makes sense to do ChIPSeq since our antibody is not very good (i.e. not realy approved) for ChIP - so we will see what works... Cheers, Tina
We did ChIP on 107 cell preparations in the EMBO paper. Sorry I can't help with your small population studies.
If you are unsure about the antibody, see if it works in IF, that is usually a good indication that it might work in ChIP.
Good luck!
Hi Jason, thanks for your response - sounds good :) We currently really depend on better antibodies to become available for this specific question. Cheers, Martina
If you can manage then you can also consider
An ultra-low-input native ChIP-seq protocol for genome-wide profiling of rare cell populations
Julie Brind’Amour1, Sheng Liu1, Matthew Hudson1, Carol Chen1, Mohammad M. Karimi1,2 & Matthew C. Lorincz1
I developed the ChIP-IT High Sensitivity assay at Active Motif with which I was able to perform ChIP on as low as 2000 cells for histone mods and 10'000 cells for TF's, however this is highly dependent on the quality of your antibody as well. If you have any questions feel free to contact me!
Hi guys, thanks very much for the helpful input. Yes, we worked with SimpleChIP and are also currently working with Active Motif - 2000 cells sounds really good! We will try the different protocols and hope to get better Abs soon since we cannot really overexpress the TF of interest in primary cells.
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