Topoisomerase II Chromatin Immunoprecipitation

Methods Mol Biol. 2018:1703:183-189. doi: 10.1007/978-1-4939-7459-7_14.

Abstract

Chromatin immunoprecipitation is a method to isolate a protein of interest coupled to DNA following cross-linking with formaldehyde and to quantify the relative abundance or occupancy of the protein at specific genomic loci. After immunoprecipitation of protein-DNA complexes protein-DNA cross-links are reversed and the DNA is extracted. Various methods exist to identify binding sites and determine relative occupancy of the protein of interest; these include quantitative PCR, probing microarrays or sequencing the isolated DNA (ChIP-seq). This chapter details the method of chromatin immunoprecipitation of TOP2 to the point of DNA extraction from the precipitated protein-DNA complexes.

Keywords: Chromatin immunoprecipitation; TOP2; Topoisomerase II.

MeSH terms

  • Binding Sites
  • Chromatin Immunoprecipitation / methods*
  • DNA / chemistry
  • DNA / metabolism*
  • DNA Topoisomerases, Type II / chemistry
  • DNA Topoisomerases, Type II / metabolism*
  • Humans
  • MCF-7 Cells
  • Poly-ADP-Ribose Binding Proteins / chemistry
  • Poly-ADP-Ribose Binding Proteins / metabolism*
  • Protein Binding
  • Sequence Analysis, DNA

Substances

  • Poly-ADP-Ribose Binding Proteins
  • DNA
  • DNA Topoisomerases, Type II
  • TOP2A protein, human
  • TOP2B protein, human