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

At 48 weeks after the transplantation surgery, ChIP of the aortic and mesenteric arteries was conducted using a ChIP-IT® kit (Active Motif, Carlsbad, CA, USA) according to the manufacturer's protocol. This involved setting up the immunoprecipitation reaction with sonicated chromatin and sequentially adding CHIP buffer and protease inhibitor cocktail to the 1.5-ml microcentrifuge tube (≥200 ng chromatin per CHIP reaction). Rabbit monoclonal anti-CREB-1-1 (ab-31387; 1:500), anti-ER-α (ab-32063; 1:200) and anti-ER-β (ab-3577; 1:500) (all from Abcam, Cambridge, MA, USA) were added and the reaction mixture was incubated overnight at 4°C. The next day, following extensive blocking in 0.5% BSA, protein G agarose beads were added to the mixture and incubated for 3 h. Following washing and the reversal of cross-links, DNA was recovered and purified. Finally, the SYBR-Green quantitative PCR was performed. Data are presented as the percentage of input.

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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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