Protocol for chromatin immunoprecipitation of histone modifications in frozen adipose tissue.

Autor:	Cayir A; EpiGen, Medical Division, Akershus University Hospital, 1478 Lørenskog, Norway., Tannæs TM; EpiGen, Medical Division, Akershus University Hospital, 1478 Lørenskog, Norway., Saeed S; Department of Clinical Molecular Biology, EpiGen, Institute of Clinical Medicine, University of Oslo, 1171 Oslo, Norway., Blüher M; Helmholtz Institute for Metabolic, Obesity and Vascular Research (HI-MAG) of the Helmholtz Zentrum München at the University of Leipzig and University Hospital, Leipzig, Germany., Böttcher Y; EpiGen, Medical Division, Akershus University Hospital, 1478 Lørenskog, Norway; Department of Clinical Molecular Biology, EpiGen, Institute of Clinical Medicine, University of Oslo, 1171 Oslo, Norway. Electronic address: yvonne.bottcher@medisin.uio.no.
Jazyk:	angličtina
Zdroj:	STAR protocols [STAR Protoc] 2024 Sep 20; Vol. 5 (3), pp. 103163. Date of Electronic Publication: 2024 Jun 27.
DOI:	10.1016/j.xpro.2024.103163
Abstrakt:	Chromatin immunoprecipitation (ChIP) combined with sequencing has revolutionized our understanding of gene regulation; however, its application to frozen adipose tissue presents unique challenges due to the high levels of lipid content. Here, we present a protocol for ChIP of histone modifications in human frozen adipose tissue. We describe steps for tissue preparation, chromatin isolation, sonication, pre-clearing of chromatin, and immunoprecipitation. We then detail procedures for elution, crosslink reversal, chromatin purification, quality control, and library synthesis. Competing Interests: Declaration of interests M.B. received personal honoraria from Amgen, AstraZeneca, Bayer, Boehringer Ingelheim, Lilly, Novo Nordisk, Novartis, and Sanofi as well as payments from Boehringer Ingelheim to the institution. (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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