RT Journal Article
T1 Chromatin immunoprecipitation improvements for the processing of small frozen pieces of adipose tissue.
A1 Castellano-Castillo, Daniel
A1 Denechaud, Pierre-Damien
A1 Moreno-Indias, Isabel
A1 Tinahones, Francisco
A1 Fajas, Lluis
A1 Queipo-Ortuño, Maria Isabel
A1 Cardona, Fernando
K1 Chromatin Immunoprecipitation
K1 Adipose Tissue
K1 Histone Modifications
K1 Active Promoters
K1 Adipocytes
K1 Diabetes Mellitus
K1 Cardiovascular Diseases
K1 Metabolic Syndrome
K1 Chromatin Shearing
AB 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.
PB Public Library of Science
YR 2018
FD 2018-02-14
LK http://hdl.handle.net/10668/12137
UL http://hdl.handle.net/10668/12137
LA en
NO Castellano-Castillo D, Denechaud PD, Moreno-Indias I, Tinahones F, Fajas L, Queipo-Ortuño MI, et al. Chromatin immunoprecipitation improvements for the processing of small frozen pieces of adipose tissue. PLoS One. 2018 Feb 14;13(2):e0192314.
DS RISalud
RD Sep 15, 2025