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

STAR Protoc

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:

Published: September 2024

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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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11261134PMC
http://dx.doi.org/10.1016/j.xpro.2024.103163DOI Listing

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