Chromatin immunoprecipitation coupled by quantitative real-time PCR as a tool for analyzing epigenetic regulation of stem cell aging

Methods Mol Biol. 2013:976:155-65. doi: 10.1007/978-1-62703-317-6_12.

Abstract

Epigenetic regulation is one of the major players in gene expression control. Histone modifications including acetylation or methylation are epigenetic mechanisms known to regulate senescence associated genes expression during adult stem cell aging. Chromatin immunoprecipitation (ChIP) assay is widely used to investigate histone modification patterns of genes of interest. For ChIP analysis, there are several conditions to be optimized empirically. In this article, not only a method of ChIP coupled by real-time quantitative PCR but also the detailed conditions is provided based on our previously published studies.

MeSH terms

  • Adult
  • Cells, Cultured
  • Cellular Senescence / physiology*
  • Chromatin Immunoprecipitation / methods*
  • Epigenesis, Genetic*
  • Humans
  • Real-Time Polymerase Chain Reaction / methods*
  • Stem Cells / cytology
  • Stem Cells / physiology*