Chromatin immunoprecipitation improvements for the processing of small frozen pieces of adipose tissue.

Details

Ressource 1Download: journal.pone.0192314.pdf (2936.74 [Ko])
State: Public
Version: Final published version
Serval ID
serval:BIB_5E840542134A
Type
Article: article from journal or magazin.
Collection
Publications
Title
Chromatin immunoprecipitation improvements for the processing of small frozen pieces of adipose tissue.
Journal
PloS one
Author(s)
Castellano-Castillo D., Denechaud P.D., Moreno-Indias I., Tinahones F., Fajas L., Queipo-Ortuño M.I., Cardona F.
ISSN
1932-6203 (Electronic)
ISSN-L
1932-6203
Publication state
Published
Issued date
2018
Peer-reviewed
Oui
Volume
13
Number
2
Pages
e0192314
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Abstract
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.

Keywords
Adipose Tissue/metabolism, Adult, Chromatin Immunoprecipitation, Female, Freezing, Genome, Human, Humans, Male, Middle Aged, Proteome, Real-Time Polymerase Chain Reaction
Pubmed
Web of science
Open Access
Yes
Create date
15/02/2018 18:25
Last modification date
20/08/2019 15:16
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