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

Methods Mol Biol. 2024:2846:35-45. doi: 10.1007/978-1-0716-4071-5_3.

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.

Keywords: Adipocyte; Adipose tissue; ChIP; Chromatin immunoprecipitation.

MeSH terms

  • Adipocytes* / cytology
  • Adipocytes* / metabolism
  • Adipose Tissue* / cytology
  • Adipose Tissue* / metabolism
  • Animals
  • Chromatin / genetics
  • Chromatin / metabolism
  • Chromatin Immunoprecipitation* / methods
  • DNA* / genetics
  • DNA* / metabolism
  • Humans
  • Protein Binding
  • Transcription Factors* / metabolism

Substances

  • DNA
  • Transcription Factors
  • Chromatin