EZ-ChIP™

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 on C2BBe1 extracts using EZ-ChIP Assay Kit (EMD Millipore, Billerica, MA) as per manufacturer’s protocol. DNA-protein complexes were immunoprecipitated with HNF-4α antibody. The purified immunoprecipitated chromatin was used for PCR analysis with NHE3 promoter-specific primers (Table 1). This primer set amplifies the NHE3 promoter region containing the consensus HNF-4α-binding direct repeat (N3-DR1 motif) of the core NHE3 promoter. PCR products were resolved on 2% agarose gel, stained with ethidium bromide, and documented using the Alpha Imager 1220 (Alpha Innotech, San Leandro, CA) gel documentation system.

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Papers

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Manufacturer protocol

Download the product protocol from Merck Millipore for EZ-ChIP™ below.

Download manufacturer protocol

Videos

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