Open Access

Easy detection of chromatin binding proteins by the histone association assay

Biological Procedures Online7:710060

https://doi.org/10.1251/bpo106

Received: 22 March 2005

Accepted: 14 April 2005

Abstract

The Histone Association Assay provides an easy approach for detecting proteins that bind chromatin in vivo. This technique is based on a chromatin immunoprecipitation protocol using histone H3-specific antibodies to precipitate bulk chromatin from crosslinked whole cell extracts. Proteins that co-precipitate with chromatin are subsequently detected by conventional SDS-PAGE and Western blot analysis. Unlike techniques that separate chromatin and nonchromatin interacting proteins by centrifugation, this method can be used to delineate whether a protein is chromatin associated regardless of its innate solubility. Moreover, the relative amount of protein bound to DNA can be ascertained under quantitative conditions. Therefore, this technique may be utilized for analyzing the chromatin association of proteins involved in diverse cellular processes.

Indexing terms

Chromatin Cross-linking Reagents Formaldehyde Histones Immunoprecipitation

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