Chromatin Immunoprecipitation (ChIP) Assay Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Just because an antibody works well in a Western blot does not always indicate it will perform well in chromatin immunoprecipitation. Unlike a Western blot that detects proteins that have been denatured, a ChIP antibody must recognize the target protein in its native state. Use ChIP-validated antibody. -Use 2-10 μg of your ChIP antibody depending on the abundance of your protein target.

Protocol tips
-Just because an antibody works well in a Western blot does not always indicate it will perform well in chromatin immunoprecipitation. Unlike a Western blot that detects proteins that have been denatured, a ChIP antibody must recognize the target protein in its native state. Use ChIP-validated antibody. -Use 2-10 μg of your ChIP antibody depending on the abundance of your protein target.

Publication protocol

The chromatin immunoprecipitation (ChIP) assay was performed using a Millipore ChIP assay kit according to the manufacturer's protocol. The antibodies used were as follows: anti-HDAC2 (Abcam), anti-H4K16ac (Millipore), and normal rabbit IgG (Santa Cruz Biotechnology); 2 μg of the specified antibody was used for each ChIP assay. qRT-PCR was performed to analyze the enrichment of sequences in DNA subjected to ChIP. The primers used for ChIP are as follows:
mBcl6-ChIP-F: 5-CTAGAGTATTAGGCACCACG-3,
mBcl6-ChIP-R: 5-CGCCCCAACATAATTGTCCC-3;
mBim-ChIP-F: 5-GGGCGGGTACATTCTGAGT-3,
mBim-ChIP-R: 5-CAGCACAGGATGTCAAAACT-3;
mp21-ChIP-F: 5-GTAAAGGCACAGGAGGTCAC-3,
mp21-ChIP-R: 5-CAGCACAGGATGTCAAAACT-3;
mp27-ChIP-F: 5-ACGCATCGCTGCTACTACAG-3,
mp27-ChIP-R: 5-CTTAGCCGACCTCACTACGG-3.

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Manufacturer protocol

Download the product protocol from Merck Millipore for Chromatin Immunoprecipitation (ChIP) Assay Kit below.

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