serval:BIB_5E840542134A
Chromatin immunoprecipitation improvements for the processing of small frozen pieces of adipose tissue.
10.1371/journal.pone.0192314
000425183500063
29444131
Castellano-Castillo
D.
author
Denechaud
P.D.
author
Moreno-Indias
I.
author
Tinahones
F.
author
Fajas
L.
author
Queipo-Ortuño
M.I.
author
Cardona
F.
author
article
2018
PloS one
1932-6203
1932-6203
journal
13
2
e0192314
Chromatin immunoprecipitation (ChIP) has gained importance to identify links between the genome and the proteome. Adipose tissue has emerged as an active tissue, which secretes a wide range of molecules that have been related to metabolic and obesity-related disorders, such as diabetes, cardiovascular failure, metabolic syndrome, or cancer. In turn, epigenetics has raised the importance in discerning the possible relationship between metabolic disorders, lifestyle and environment. However, ChIP application in human adipose tissue is limited by several factors, such as sample size, frozen sample availability, high lipid content and cellular composition of the tissue. Here, we optimize the standard protocol of ChIP for small pieces of frozen human adipose tissue. In addition, we test ChIP for the histone mark H3K4m3, which is related to active promoters, and validate the performance of the ChIP by analyzing gene promoters for factors usually studied in adipose tissue using qPCR. Our improvements result in a higher performance in chromatin shearing and DNA recovery of adipocytes from the tissue, which may be useful for ChIP-qPCR or ChIP-seq analysis.
Adipose Tissue/metabolism
Adult
Chromatin Immunoprecipitation
Female
Freezing
Genome, Human
Humans
Male
Middle Aged
Proteome
Real-Time Polymerase Chain Reaction
eng
60_published
true
peer-reviewed
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
University of Lausanne
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