Chromatin ip assay kit

Manufactured by Merck Group

The Chromatin IP Assay Kit is a laboratory tool designed to study protein-DNA interactions. It provides a method for isolating and analyzing specific DNA sequences associated with DNA-binding proteins. The kit includes reagents and protocols for chromatin immunoprecipitation (ChIP) experiments, which are used to investigate gene regulation and epigenetic mechanisms.

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Lab products found in correlation

Ab5079, by Abcam (1 mentions) Ultrasonic cell disruptor, by Emerson (1 mentions) Dna extraction kit, by Qiagen (1 mentions) E220 focused ultrasonicator, by Covaris (1 mentions)

2 protocols using chromatin ip assay kit

ChIP Assay of FOXC1 Binding

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RAW264.7 cells were crosslinked with 1% (v/v) formaldehyde and were subjected to ChIP assays using Chromatin IP Assay Kit (Millipore). Briefly, cells were lysed and then sonicated to obtain DNA fragments (200–1000 bp in size) using an ultrasonic cell disruptor (Branson, Danbury, CT, USA). The soluble chromatin was then immunoprecipitated with anti-FOXC1 antibody (ab5079; Abcam; 1:50) overnight at 4 °C with rotation and then supplemented with protein A agarose/salmon sperm DNA for 1 h at 4 °C. Reversing the cross-links was carried out for 4 h at 65 °C. DNA was purified with a DNA extraction kit (Qiagen). Finally, the purified DNA was analyzed by qPCR. Primers for qPCR are listed in Table S1. P1–P3 denote the three promoter regions of Foxc1 gene analyzed in ChIP assays, and P4 is at the distal region²⁸ .

He T., Shang J., Gao C., Guan X., Chen Y., Zhu L., Zhang L., Zhang C., Zhang J, & Pang T. (2020). A novel SIRT6 activator ameliorates neuroinflammation and ischemic brain injury via EZH2/FOXC1 axis. Acta Pharmaceutica Sinica. B, 11(3), 708-726.

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ChIP-qPCR for Transcription Factors

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Chromatin immunoprecipitation assays were performed with the chromatin IP Assay kit (Millipore, Temecula, Calif). Briefly, cells were fixed in a formaldehyde solution and then resuspended in SDS lysis buffer. Lysates were sonicated with an E220 focused-ultrasonicator (Covaris, Woburn, Mass) to a desired fragment size distribution of 100 to 200 bp. Immunoprecipitation reactions were performed with anti-RORgt (eBioscience) or anti-nuclear receptor subfamily 2 group F member 6 (Nr2f6; GeneTex, Irvine, Calif) antibodies. Chromatin immunoprecipitates were quantified by using the realtime PCR system.

Tamehiro N., Nishida K., Sugita Y., Hayakawa K., Oda H., Nitta T., Nakano M., Nishioka A., Yanobu-Takanashi R., Goto M., Okamura T., Adachi R., Kondo K., Morita A, & Suzuki H. (2019). Ras homolog gene family H (RhoH) deficiency induces psoriasis-like chronic dermatitis by promoting T_H17 cell polarization. The Journal of allergy and clinical immunology, 143(5).

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