Chromatin Immunoprecipitation in Skeletal Myoblasts

Methods Mol Biol. 2019:1889:43-54. doi: 10.1007/978-1-4939-8897-6_4.

Abstract

Chromatin immunoprecipitation (ChIP) is a powerful and sensitive technique that is widely used to study DNA-protein interactions. It enables an unbiased genome-wide analysis of transcriptional changes during several biological processes including cellular differentiation. Here, we describe a step-by-step protocol to identify histone modifications, transcription factor, and co-factor binding to chromatin in skeletal myoblasts. We discuss critical steps during cell harvesting, sonication, and immunoprecipitation and provide notes to evade common pitfalls.

Keywords: Chromatin modifiers; Epigenetics; Histone modifications; Myoblasts; Transcription.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatin / genetics
  • Chromatin / metabolism
  • Chromatin Immunoprecipitation*
  • Data Analysis
  • High-Throughput Nucleotide Sequencing
  • Histones / metabolism
  • Myoblasts, Skeletal / metabolism*
  • Real-Time Polymerase Chain Reaction

Substances

  • Chromatin
  • Histones