Detection of E2F-DNA Complexes Using Chromatin Immunoprecipitation Assays. Academic Article uri icon

Overview

abstract

  • Chromatin immunoprecipitation (ChIP), originally developed by John T. Lis and David Gilmour in 1984, has been useful to detect DNA sequences where protein(s) of interest bind. ChIP is comprised of several steps: (1) cross-linking of proteins to target DNA sequences, (2) breaking genomic DNA into 300-1000 bp pieces by sonication or nuclease digestion, (3) immunoprecipitation of protein bound to target DNA with an antibody, (4) reverse cross-linking between target DNA and the bound protein to liberate the DNA fragments, and (5) amplification of target DNA fragment by PCR. Since then, the technology has evolved significantly to allow not only amplifying target sequences by PCR, but also sequencing all DNA fragment bound to a target protein, using a variant of the approach called the ChIP-seq technique (1). Another variation, the ChIP-on-ChIP, allows the detection of protein complexes bound to specific DNA sequences (2).

publication date

  • January 1, 2018

Research

keywords

  • Chromatin
  • Chromatin Immunoprecipitation
  • DNA
  • E2F Transcription Factors
  • High-Throughput Nucleotide Sequencing
  • Polymerase Chain Reaction

Identity

PubMed Central ID

  • PMC6070307

Scopus Document Identifier

  • 85042446576

Digital Object Identifier (DOI)

  • 10.1007/978-1-4939-7565-5_13

PubMed ID

  • 29468550

Additional Document Info

volume

  • 1726