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

EZ ChIP™ Chromatin Immunoprecipitation Kit (Millipore, Bedford, MA, USA) and the EpiQuik Tissue Acetyl-Histone H3 and H4 CHIP Kit (Epigentek Group Inc., NY, USA) were used to perform ChIP assays. In brief, cells were crosslinked with 4% formaldehyde for 10 min followed by cell collection and sonication with a predetermined power to yield genomic DNA fragments of 300–1000 bp long. The chromatin was immunoprecipitated using an anti-acetyl-histone H3 or anti-acetyl-histone H4 antibody, or anti-FOXO3 antibody. Normal mouse IgG or normal rabbit IgG were used as the negative control. qRT-PCR was conducted using KAPA SYBR FAST qPCR Kit (Kapa Biosystems, USA). PCR products were analyzed by agarose gel electrophoresis.

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Manufacturer protocol

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

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