Biotechniques. 2004 Dec;37(6):961-9. doi: 10.2144/04376RV01.

Chromatin immunoprecipitation assay.

BioTechniques

Partha M Das, Kavitha Ramachandran, Jane vanWert, Rakesh Singal

PMID: 15597545 DOI: 10.2144/04376RV01

Abstract

Association between proteins and DNA is crucial for many vital cellular functions such as gene transcription, DNA replication and recombination, repair, segregation, chromosomal stability, cell cycle progression, and epigenetic silencing. It is important to know the genomic targets of DNA-binding proteins and the mechanisms by which they control and guide gene regulation pathways and cellular proliferation. Chromatin immunoprecipitation (ChIP) is an important technique in the study of protein-gene interactions. Using ChIP, DNA-protein interactions are studied within the context of the cell. The basic steps in this technique are fixation, sonication, immunoprecipitation, and analysis of the immunoprecipitated DNA. Although ChIP is a very versatile tool, the procedure requires the optimization of reaction conditions. Several modifications to the original ChIP technique have been published to improve the success and to enhance the utility of this procedure. This review addresses the critical parameters and the variants of ChiP as well as the different analytical tools that can be combined with ChIP to enable better understanding of DNA-protein interactions in vivo.

Substances

• DNA-Binding Proteins
• DNA

MeSH terms

• Chromatin Immunoprecipitation / instrumentation
• Chromatin Immunoprecipitation / methods
• Chromatin Immunoprecipitation / trends
• DNA / analysis
• DNA / metabolism
• DNA-Binding Proteins / analysis
• DNA-Binding Proteins / metabolism

Publication Types

• Journal Article
• Research Support, U.S. Gov't, Non-P.H.S.
• Research Support, U.S. Gov't, P.H.S.
• Review