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

ChIP was performed using the ChIP-IT Express Enzymatic kit (Active Motif, Carlsbad, CA) according to the manufacturer’s instructions. Immunoprecipitation of 25 μg DNA was performed overnight at 4°C using anti-p65 antibodies (cat. no. 8242; Cell Signaling Technology). DNA isolated from an aliquot of the total nuclear extract was used as a loading control for the PCR (input control). PCR was performed with total DNA and immunoprecipitated DNA using the following promoter-specific primers: IL1β promoter-binding site for NF-κB: 5’-CAGACAGGGAGGGCTATTGG-3’ (forward), 5’-GGTTCATGGAAGGGCAAGGA-3’ (reverse) (297 bp), and TNF promoter binding site for NF-κB: 5’-GATGGGGAGTGTGAGGGGTA-3’ (forward), 5’-CAGCGGAAAACTTCCTTGGTG-3’ (reverse) (159 bp). The PCR products (10 μL) were subjected to electrophoresis on a 2% agarose gel.

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Papers

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Manufacturer protocol

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

Download manufacturer protocol

Videos

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