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
收藏
页数：20
相关论文
共 50 条
  • [31] Regulation of chromatin by histone modifications
    Bannister, Andrew J.
    Kouzarides, Tony
    CELL RESEARCH, 2011, 21 (03) : 381 - 395
  • [32] Regulation of chromatin by histone modifications
    Andrew J Bannister
    Tony Kouzarides
    Cell Research, 2011, 21 : 381 - 395
  • [33] HISTONE CONFORMATIONS, HISTONE MODIFICATIONS, AND CHROMATIN STRUCTURE
    BRADBURY, EM
    DIFFERENTIATION, 1979, 13 (01) : 37 - 39
  • [34] HISTONE INTERACTIONS, HISTONE MODIFICATIONS AND CHROMATIN STRUCTURE
    BRADBURY, EM
    PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY OF LONDON SERIES B-BIOLOGICAL SCIENCES, 1978, 283 (997) : 291 - 293
  • [35] Protocol for the fast chromatin immunoprecipitation (ChIP) method
    Nelson, Joel D.
    Denisenko, Oleg
    Bomsztyk, Karol
    NATURE PROTOCOLS, 2006, 1 (01) : 179 - 185
  • [36] Protocol for the fast chromatin immunoprecipitation (ChIP) method
    Joel D Nelson
    Oleg Denisenko
    Karol Bomsztyk
    Nature Protocols, 2006, 1 : 179 - 185
  • [37] An optimised chromatin immunoprecipitation (ChIP) method for starchy leaves of Nicotiana benthamiana to study histone modifications of an allotetraploid plant
    Buddhini Ranawaka
    Milos Tanurdzic
    Peter Waterhouse
    Fatima Naim
    Molecular Biology Reports, 2020, 47 : 9499 - 9509
  • [38] Genome-wide Analysis of Histone Modifications Distribution using the Chromatin Immunoprecipitation Sequencing Method in Magnaporthe oryzae
    Wu, Zechi
    Sun, Wanyu
    Zhou, Sida
    Zhang, Li
    Zhao, Xinyu
    Xu, Yang
    Wang, Weixiang
    JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2021, (172):
  • [39] Chromatin immunoprecipitation (ChIP) method for non-model fruit flies (Diptera: Tephritidae) and evidence of histone modifications
    Nagalingam, Kumaran
    Lorenc, Michel T.
    Manoli, Sahana
    Cameron, Stephen L.
    Clarke, Anthony R.
    Dudley, Kevin J.
    PLOS ONE, 2018, 13 (03):
  • [40] An optimised chromatin immunoprecipitation (ChIP) method for starchy leaves of Nicotiana benthamiana to study histone modifications of an allotetraploid plant
    Ranawaka, Buddhini
    Tanurdzic, Milos
    Waterhouse, Peter
    Naim, Fatima
    MOLECULAR BIOLOGY REPORTS, 2020, 47 (12) : 9499 - 9509
12345