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

Chromatin immunoprecipitation (ChIP) assays were performed using the ChIP Assay Kit (Millipore, Billerica, USA). Rat liver tissues were homogenised and fixed with 1% formaldehyde. DNA was sheared to 200–1000 bp fragments using sonication. Chromatin was incubated and precipitated with antibodies against FXR, histone deacetylase 3 (HDAC3; Epitomics), or IgG (Boster, Wuhan, China). Primers used to amplify the binding site of FXR on the promoter region of MRP2 gene for qPCR were designed as follows: sense- 5’-agagtgagttccaggacagt-3’; antisense- 5’-ccctgtagtgtgaccaaata-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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