Chromatin Immunoprecipitation in Adipose Tissue and Adipocytes: How to Proceed and Optimize the Protocol for Transcription Factor DNA Binding.

Adipocytes / metabolism Adipose Tissue / metabolism Chromatin Immunoprecipitation / methods DNA / metabolism Transcription Factors / metabolism Humans Animals Protein Binding Chromatin / metabolism
Adipocyte Adipose tissue ChIP Chromatin immunoprecipitation

Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2024
Historique:
medline: 14 8 2024
pubmed: 14 8 2024
entrez: 14 8 2024
Statut: ppublish

Résumé

Chromatin immunoprecipitation (ChIP) coupled to qPCR or sequencing is a crucial experiment to determine direct transcriptional regulation under the control of specific transcriptional factors or co-regulators at loci-specific or pan-genomic levels.Here we provide a reliable method for processing ChIP from adipocytes or frozen adipose tissue collection, isolation of nuclei, cross-linking of protein-DNA complexes, chromatin shearing, immunoprecipitation, and DNA purification. We also discuss critical steps for optimizing the experiment to perform a successful ChIP in lipid-rich cells/tissues.

Identifiants

DOI: 10.1007/978-1-0716-4071-5_3 PMID: 39141228
pubmed: 39141228
doi: 10.1007/978-1-0716-4071-5_3
doi:

Substances chimiques

DNA 9007-49-2
Transcription Factors 0
Chromatin 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

35-45

Informations de copyright

© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Références

Mir AA, Dyar KA, Greulich F et al (2019) In vivo ChIP-Seq of nuclear receptors: a rough guide to transform frozen tissues into high-confidence genome-wide binding profiles. Methods Mol Biol 1966:39–70
doi: 10.1007/978-1-4939-9195-2_5 pubmed: 31041738
Giroud M, Tsokanos FF, Caratti G et al (2021) HAND2 is a novel obesity-linked adipogenic transcription factor regulated by glucocorticoid signalling. Diabetologia 64(8):1850–1865
doi: 10.1007/s00125-021-05470-y pubmed: 34014371 pmcid: 8245394
van Eenige R, In Het Panhuis W, Schonke M et al (2022) Angiopoietin-like 4 governs diurnal lipoprotein lipase activity in brown adipose tissue. Mol Metab 60:101497
doi: 10.1016/j.molmet.2022.101497 pubmed: 35413480 pmcid: 9048098

Auteurs

Céline Jouffe (C)

Institute for Diabetes and Cancer, Helmholtz Zentrum Muenchen, Neuherberg, Germany. celine.jouffe@helmholtz-munich.de.
Institute for Diabetes and Endocrinology, Helmholtz Zentrum Muenchen, Neuherberg, Germany. celine.jouffe@helmholtz-munich.de.

Kenneth A Dyar (KA)

Institute for Diabetes and Cancer, Helmholtz Zentrum Muenchen, Neuherberg, Germany.
German Center for Diabetes Research (DZD), Neuherberg, Germany.

Nina Henriette Uhlenhaut (NH)

Institute for Diabetes and Endocrinology, Helmholtz Zentrum Muenchen, Neuherberg, Germany.
Metabolic Programming, TUM School of Life Sciences Weihenstephan, & ZIEL-Institute for Food & Health, Freising, Germany.

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Classifications MeSH

questionsmedicales.fr › Cellules › Cellules du tissu conjonctif › Adipocytes › Chromatin Immunoprecipitation in Adipose...
questionsmedicales.fr › Tissus › Tissu conjonctif › Tissu adipeux › Chromatin Immunoprecipitation in Adipose...
questionsmedicales.fr › Techniques d'investigation › Techniques immunologiques › Immunoprécipitation › Immunoprécipitation de la chromatine › Chromatin Immunoprecipitation in Adipose...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ADN › Chromatin Immunoprecipitation in Adipose...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Facteurs de transcription › Chromatin Immunoprecipitation in Adipose...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Primates › Haplorhini › Catarrhini › Hominidae › Humains › Chromatin Immunoprecipitation in Adipose...
questionsmedicales.fr › Eucaryotes › Animaux › Chromatin Immunoprecipitation in Adipose...
questionsmedicales.fr › Métabolisme › Liaison aux protéines › Chromatin Immunoprecipitation in Adipose...
questionsmedicales.fr › Phénomènes génétiques › Structures génétiques › Structures de chromosome › Chromatine › Chromatin Immunoprecipitation in Adipose...