EZ-Magna ChIP™ A - 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 assays were performed in Hepa1–6 cells using EZ-Magna ChIPTM A (Cat. #17–408, Millipore) following the standard protocol. The protein-DNA complexes were incubated with 5 µg of anti-Runx2 antibody (CST, #8486) or 5 µg of normal rabbit IgG (provided in the EZ-Magna ChIPTM A kit) as the immunoprecipitating antibody. Purified DNA was subjected to RT-PCR (ExTaq, Takara) and qPCR (SYBR Green PCR Master Mix, TaKaRa) analyses using Runx2 ChIP primers. The positive control was incubated with 5 µg anti-acetyl Histone H3 (provided by EZ-Magna ChIPTM A) and purified DNA was detected by RT-PCR using Gapdh ChIP primers.

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Manufacturer protocol

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

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