《蛋白质-核酸相互作用技术》相关实验技术-ChIP-a tool for studying histone acetylation and transcription factor binding.pdfVIP

Methods 31 (2003) 67–75 /locate/ymeth Chromatin immunoprecipitation: a tool for studying histone acetylation and transcription factor binding Virginia A. Spencer, Jian-Min Sun, Lin Li, and James R. Davie* Manitoba Institute of Cell Biology, University of Manitoba, Winnipeg, Manitoba, Canada R3E 0V9 Accepted 19 February 2003 Abstract The function of a protein in gene expression can often be explained, in part, by the location of that protein along a specific gene sequence. In recent years, the chromatin immunoprecipitation (ChIP) assay has been developed to study the association of proteins located within 2 AA of DNA such as transcription factors and modified histones. Numerous important findings have been published using the ChIP assay and many questions about transcription have been answered. In this article, we present the ChIP assay currently used in our lab and discuss the various ways to optimize this assay for ones own use. 2003 Published by Elsevier Science (USA). Keywords: Chromatin immunoprecipitation; Histone; Transcription factor; Formaldehyde; Cisplatin 1. Introduction Initially, the ChIP assay was used to study the asso- ciation of hyperacetylated histones with specific DNA The chromatin immunoprecipitation (ChIP) assay sequences to further understand the role of histone has become a very popular technique for fine-mapping acetylation in transcription [1]. However, more recent the location of modified histone