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ChIP Chromatin Immunoprecipitation Forum Chromatin Immunoprecipitation Forum. Discuss, chat about, and post questions about the ChIP assay, in the ChIP forum. Includes troubleshooting, protocols, buffer preparation, and more.


ChIP with 100% confluent cells

ChIP with 100% confluent cells - ChIP Chromatin Immunoprecipitation Forum

ChIP with 100% confluent cells - Chromatin Immunoprecipitation Forum. Discuss, chat about, and post questions about the ChIP assay, in the ChIP forum. Includes troubleshooting, protocols, buffer preparation, and more.


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Old 06-17-2010, 06:06 PM
Pipette Filler
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Default ChIP with 100% confluent cells



Hi all,

I'm using NHEK cells and growing them to 100% confluence because they react to my treatment best that way. The problem is I'm having trouble lysing and homogenizing the cells, they get stuck in clumps and my nuclei yield is very low. It doesn't matter if I homogenize 5 times or 100 times, I still get the same results: low input. Any suggestions on what I should do?

I'm currently using the Active Motif enzymatic ChIP-it express kit.

Thanks in advance for any help
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Old 05-16-2012, 09:11 PM
Pipette Filler
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Default Re: ChIP with 100% confluent cells

You can opt for sonication instead of homogenisation and then maybe use the enzyme kit, I had the same problem but it helped.
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