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

ChIP assays were performed using EZ-CHIP Kit according to the manufacturer’s instruction (Millipore, USA). Briefly, cells were cross-linked using 1% formaldehyde. After washing with PBS, the cells were resuspended in 1 mL of SDS lysis buffer containing protease inhibitor cocktail. DNA was sheared to small fragments (200–300 bp) by sonication. The sheared chromatin was immunoprecipitated with different antibodies (Additional file 2) overnight at 4 °C. DNA from immunoprecipitation and the input samples was analyzed using SYBR Green qPCR Master Mix (Biotool, USA).

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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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