MAGnify™ Chromatin Immunoprecipitation System

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-The 1.25 M glycine must be at room temperature before use. -The Lysis Buffer must be at room temperature and fully resuspended before use. Vortex briefly to resuspend.	-For each ChIP reaction, use 10,000–300,000 cells or 0.167–5 mg of tissue. -1–10 μg of antibody is a typical starting range. - 10-minute crosslinking step using formaldehyde at a 1% final concentration. -Keep the cell lysate cooled on ice during sonication.

Upstream tips
-The 1.25 M glycine must be at room temperature before use. -The Lysis Buffer must be at room temperature and fully resuspended before use. Vortex briefly to resuspend.
Protocol tips
-For each ChIP reaction, use 10,000–300,000 cells or 0.167–5 mg of tissue. -1–10 μg of antibody is a typical starting range. - 10-minute crosslinking step using formaldehyde at a 1% final concentration. -Keep the cell lysate cooled on ice during sonication.

Publication protocol

Chromatin immunoprecipitation (ChIP) assay was performed using a MAGnify™ ChIP kit (Invitrogen) following the manufacturer’s protocol. Immune complexes from monolayer culture and spheres derived from SCC4were obtained using 5 µg of JMJD6 antibody (H-7; Santa Cruz Biotech). Then, genomic DNA was isolated from the complexes and subjected to qPCR using the following IL4 promoter primers: sense, 5′-GCCTGTTATTCTGCCTCTATGC-3′; antisense, 5′-TGGAAACTGTCCTGTCATGG-3′.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for MAGnify™ Chromatin Immunoprecipitation System below.

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