Chromatin Immunoprecipitation from Formaldehyde Cross-Linked Olfactory Sensory Neurons

Methods Mol Biol. 2023:2710:71-82. doi: 10.1007/978-1-0716-3425-7_6.

Abstract

Chromatin immunoprecipitation (ChIP) allows a researcher to determine the genomic occupancy of nuclear proteins, providing insight into the roles of transcription factors, chromatin modifiers, histone modifications, and other factors bound to DNA. Protein-DNA interactions are first fixed in vivo by chemical cross-linking, and then a target protein is captured together with any associated DNA by an antibody mediated pull-down. The co-immunoprecipitated DNA can then be assayed by quantitative PCR or deep sequencing. Here, we demonstrate this technique using murine olfactory sensory neurons (OSNs) purified using fluorescence-activated cell sorting (FACS) and antibodies for the ubiquitous chromatin protein CTCF.

Keywords: CTCF; Chromatin; Cross-link; Formaldehyde; Immunoprecipitation; Olfactory sensory neuron; Sonication.

MeSH terms

  • Animals
  • Antibodies
  • Chromatin / genetics
  • Chromatin Immunoprecipitation
  • Formaldehyde
  • Mice
  • Olfactory Receptor Neurons*

Substances

  • Formaldehyde
  • Antibodies
  • Chromatin