ChIP-IT® Express Chromatin Immunoprecipitation Kits

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Do not re-freeze the Protein G magnetic beads, the beads should be stored at 4C.	-Always use ChIP validated antibody. -Optimal use of 1-3 ug of antibody.	-The diluted Proteinase K stop solution can not be stored.

Upstream tips
-Do not re-freeze the Protein G magnetic beads, the beads should be stored at 4C.
Protocol tips
-Always use ChIP validated antibody. -Optimal use of 1-3 ug of antibody.
Downstream tips
-The diluted Proteinase K stop solution can not be stored.

Publication protocol

A ChIP assay was performed using the ChIP IT Express Enzymatic Kit (Active Motif, Carlsbad, CA), according to the manufacturer's instructions. In brief, H4IIEC3 hepatocytes were grown in 15-cm dishes and transfected with 30.4 μg of pcDNA3.1(+) mCherry-mature SREBP-1c plasmid vectors per dish together with 91.2 μl of FuGENE6 (Promega). Forty-eight hours later, cells were treated with 0.25 mm EPA for 24 h before being fixed and homogenized. Following centrifugation, the supernatant was used for chromatin samples. Chromatin samples were incubated with protein G-coated magnetic beads and RFP antibodies (Medical & Biological Laboratories Co., Ltd., Nagoya, Japan) overnight at 4 °C. Following washing and elution, a reaction solution was used as the template for PCR. PCR primers were set for amplification of the Mut-DΔ2 region of the SELENOP promoter, as follows: forward, 5′-AACATTCTTCTCGTCGCGGCAACCA-3′; and reverse, 5′-AGATCCACAAAGCCACAGGCTGACA-3′.

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Manufacturer protocol

Download the product protocol from Active Motif for ChIP-IT® Express Chromatin Immunoprecipitation Kits below.

Download manufacturer protocol

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