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.
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页数：20
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