Chromatin Immunoprecipitation in Adipose Tissue and Adipocytes: How to Proceed and Optimize the Protocol for Transcription Factor DNA Binding.
Methods Mol Biol
; 2846: 35-45, 2024.
Article
em En
| MEDLINE
| ID: mdl-39141228
ABSTRACT
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.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Fatores de Transcrição
/
DNA
/
Tecido Adiposo
/
Adipócitos
/
Imunoprecipitação da Cromatina
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Methods Mol Biol
Ano de publicação:
2024
Tipo de documento:
Article