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

Chromatin was extracted from 20 million Hepa 1.6 or NIH3T3 cells with ChIP-IT Express Enzymatic kit (Active Motif), according to the manufacturer’s recommendations. Every ChIP reaction was performed with 5 μg of mouse anti-HNF4α antibodies (H1415, Life Technologies). Immunoprecipitated DNA and Input DNA were purified with Chromatin IP DNA Purification Kit (Active Motif). It was subsequently analyzed by qPCR to measure the relative enrichment of the fragments of interest in the total input of ChIP DNA fragments. The negative control ChIP primers were designed to amplify fragments downstream of the Agxt2 predicted transcription start sites. The positive control primers were designed to amplify the region of hepatocyte nuclear factor 1 alpha (Hnf1a) promoter, which has been shown previously to comprise an active HNF4A binding site28. Murine fibroblast cell line NIH 3T3 was used as an additional negative control. ChIP-IT Control qPCR Kit for Mouse (Active Motif) was used as an additional control.

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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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