Protocol for chromatin immunoprecipitation of histone modifications in frozen adipose tissue

被引：0

作者：

Cayir, Akin ^{[1
]}

Tannaes, Tone M. ^{[1
]}

Saeed, Sadia ^{[2
]}

Blueher, Matthias ^{[3
]}

Boettcher, Yvonne ^{[1
,2
,4
]}

机构：

[1] Akershus Univ Hosp, Med Div, EpiGen, N-1478 Lorenskog, Norway

[2] Univ Oslo, Inst Clin Med, Dept Clin Mol Biol, EpiGen, N-1171 Oslo, Norway

[3] Univ Leipzig, Helmholtz Inst Metab Obes & Vasc Res HI MAG, Helmholtz Zentrum Munchen, Leipzig, Germany

[4] Univ Hosp Leipzig, Leipzig, Germany

来源：

STAR PROTOCOLS | 2024年 / 5卷 / 03期

关键词：

Chromatin immunoprecipitation (ChIP); Health Sciences;

D O I：

10.1016/j.xpro.2024.103163

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页数：20