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 was performed according to the manufacturer’s protocol. Opossum Kidney cells were transfected using lipofectamin with TRPC6 promoter construct, and untransfected podocytes were treated with pioglitazone, after which a ChIP was performed (Magna ChIP A Chromatin Immunoprecipitation Kit; Upstate Millipore) using an antibody directed against PPAR-γ (Santa Cruz), an appropriate isotype control, and a positive control, an antibody directed against histone H3. The enrichment of the TRPC6 promoter compared with the isotype control was evaluated using real-time qPCR with specific primers corresponding to the TRPC6 promoter region. Subsequently, an immunoblot of all samples was done to determine PPAR-γ (Santa Cruz) expression in the input and immunoprecipitated samples.

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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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