2. Academic Validation
  3. Chromatin Immunoprecipitation (ChIP) to Study DNA-Protein Interactions

Chromatin Immunoprecipitation (ChIP) to Study DNA-Protein Interactions

  • Methods Mol Biol. 2021:2261:323-343. doi: 10.1007/978-1-0716-1186-9_20.
Eliza C Small 1,  Danielle N Maryanski 2,  Keli L Rodriguez 2,  Kevin J Harvey 3,  Michael-C Keogh 2,  Andrea L Johnstone 2
Affiliations

Affiliations

  • 1 Thermo Fisher Scientific, Rockford, IL, USA. [email protected].
  • 2 EpiCypher Inc., Durham, NC, USA.
  • 3 Thermo Fisher Scientific, Carlsbad, CA, USA.
PMID: 33420999 DOI: 10.1007/978-1-0716-1186-9_20
Abstract

Chromatin immunoprecipitation (ChIP) is a method used to examine the genomic localization of a target of interest (e.g., proteins, protein posttranslational modifications, or DNA elements). As ChIP provides a snapshot of in vivo DNA-protein interactions, it lends insight to the mechanisms of gene expression and genome regulation. This chapter provides a detailed protocol focused on native-ChIP (N-ChIP), a robust approach to profile stable DNA-protein interactions. We also describe best practices for ChIP , including defined controls to ensure specific and efficient target enrichment and methods for data normalization.

Keywords

Antibody specificity; ChIP; ChIP normalization; Chromatin immunoprecipitation; Histone PTMs; Recombinant nucleosomes; Semisynthetic nucleosomes; Spike-in controls.

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