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ChIP with low cell number

chromatin immunoprecipitation cell number quantitation % Input

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#1 hotstuffdb22

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Posted 21 July 2013 - 12:57 PM

Hi everyone! I want to isolate fresh B-cells from mice and then do ChIP immediately, as opposed to growing the cells in culture for several days. I believe I'll have at least 3 million cells after isolation and I want to do ChIP using 5 different antibodies, including IgG. My final calculation of qPCR data will be based on percentage of input, all to avoid quantitation by fluorimetry (and more loss of material). My question is whether 3 million cells will be sufficient for 5 different antibodies (600,000 cells each). Has anyone done this before? Thanks!

#2 chabraha

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Posted 22 July 2013 - 08:11 AM

after you get the shearing conditions down this will easily work..................use the mu-ChIP protocols out there
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#3 hotstuffdb22

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Posted 23 July 2013 - 02:29 PM

I plan to start with 3 million cells and will use normal shearing conditions that I use for >5 million cells.

Here are the antibodies I will use:
H3K9Ac
H3K9me2
H3K4me3
H3K27Me3
H3K4Me2





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