EZ-Magna ChIP™ A/G Chromatin Immunoprecipitation 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

An EZ-Magna ChIP kit (EMD Millipore, Billerica, MA, USA) was used for the ChIP experiment, according to the manufacturer's protocol. OSCC cells were treated with formaldehyde and incubated for 10 min at room temperature to generate DNA-protein cross-links. Cell lysates were then sonicated to generate chromatin fragments of 200-300 bp and immunoprecipitated for 1 h at room temperature with antibodies, including anti-H3K27ac (Abcam; 1:100; cat. no. ab4729), anti-CBP (Abcam; 1:100; cat. no. ab2832) or normal mouse IgG polyclonal antibody (EMD Millipore; 1:100; cat. no. 12-371). Precipitated chromatin DNA was recovered and analyzed by RT-qPCR as aforementioned.

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Manufacturer protocol

Download the product protocol from Merck Millipore for EZ-Magna ChIP™ A/G Chromatin Immunoprecipitation Kit below.

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