title: Chromatin immunoprecipitation for determining the association of proteins with specific genomic sequences in vivo.
abstract: Chromatin immunoprecipitation (ChIP) is a powerful and widely applied technique for detecting the association of individual proteins with specific genomic regions in vivo. Live cells are treated with formaldehyde to generate protein-protein and protein-DNA cross-links between molecules that are in close proximity on the chromatin template in vivo. DNA sequences that cross-link with a given protein are selectively enriched, and reversal of the formaldehyde cross-linking permits recovery and quantitative analysis of the immunoprecipitated DNA. As formaldehyde inactivates cellular enzymes essentially immediately upon addition to cells, ChIP provides snapshots of protein-protein and protein-DNA interactions at a particular time point, and hence is useful for kinetic analysis of events occurring on chromosomal sequences in vivo. In addition, ChIP can be combined with microarray technology to identify the location of specific proteins on a genome-wide basis. in this unit describes the ChIP procedure for Saccharomyces cerevisiae; describes the corresponding steps for mammalian cells.
authors.affiliation: University of Southern California, Los Angeles, California, USA.
authors.forename: Oscar
authors.initials: O
authors.lastname: Aparicio
authors.forename: Joseph V
authors.initials: JV
authors.lastname: Geisberg
authors.forename: Edward
authors.initials: E
authors.lastname: Sekinger
authors.forename: Annie
authors.initials: A
authors.lastname: Yang
authors.forename: Zarmik
authors.initials: Z
authors.lastname: Moqtaderi
authors.forename: Kevin
authors.initials: K
authors.lastname: Struhl
mesh_terms: D000818:Animals; D001483:Base Sequence; D002477:Cells / Q000737:chemistry / Q000378:metabolism; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D003432:Cross-Linking Reagents / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000378:metabolism*; D016678:Genome; D006801:Humans; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism
chemical_list: D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins