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

ChIP assay was performed to confirm the interaction between EZH2 and FBXO32 using the Magna ChIP™ A/G Chromatin Immunoprecipitation Kit (Millipore, Bellerica, MA, USA). Briefly, AGS/5-FU and SGC-7901/5-FU cells were lysed in a complete ChIP lysis buffer. Then, cell lysates were incubated with protein A/G magnetic beads and antibodies against EZH2 (Active Motif, Carlsbad, CA, USA), H3K27me3 (Millipore) or IgG (Millipore). Finally, qRT-PCR analysis was performed to determine the enrichment of FBXO32 in the immunoprecipitated DNA. The primers used were as follows: forward, 5′-GCAGCTATACGAGGTAGGACTGGGG-3′; reverse, 5′-TTGCAGTGAGCTGAGATCGCGC-3′.

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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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