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

A chromatin immunoprecipitation (ChIP) assay kit (EMD Millipore, Billerica, MA, USA) was used. In brief, lysates from dispersed rat islets were fixed with formaldehyde. DNA was sheared to fragments at 200–1000 bp using sonication. Chromatins were incubated and precipitated with Abs against CREB or IgG (both from Cell Signaling Technology). DNA pellets were analyzed by real-time qPCR by using primers directed to the Tph1 promoter (forward: 5′-AACCCACCAATGTGGAGAGC-3′; reverse: 5′-TACAAGGACATGAACCGCCC-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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