title: Micro chromatin immunoprecipitation (μChIP) from early mammalian embryos.
abstract: Chromatin immunoprecipitation (ChIP) is a powerful method for mapping protein-DNA interactions in vivo. Genomic localization of histone modifications, transcription factors, and other regulatory proteins can be revealed by ChIP. However, conventional ChIP protocols require the use of large numbers of cells, which prevents the application of ChIP to rare cell types. We have developed ChIP assays suited for the immunoprecipitation of histone proteins or transcription factors from small cell numbers. Here we describe a rapid, yet sensitive micro (μ)ChIP protocol producing high signal to noise ratio output, suitable for as few as 100 cells. This chapter provides a detailed protocol for μChIP from early mammalian embryos, also suitable for any sample of limited numbers of cells. Minor modifications of this optimized high signal to noise ChIP protocol make it a reliable tool for the use with any cell number (100-10(7)).
authors.affiliation: Institute of Clinical Medicine, Division of diagnostics and intervention, Department of Microbiology, Oslo university hospital, Rikshospitalet, NO-0027, Oslo, Norway, j.a.dahl@medisin.uio.no.
authors.forename: John Arne
authors.initials: JA
authors.lastname: Dahl
authors.forename: Arne
authors.initials: A
authors.lastname: Klungland
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D002417:Cattle; D002843:Chromatin / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D003432:Cross-Linking Reagents / Q000737:chemistry; D004247:DNA / Q000302:isolation & purification; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D006657:Histones / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D014157:Transcription Factors / Q000235:genetics
chemical_list: D002843:Chromatin; D003432:Cross-Linking Reagents; D006657:Histones; D014157:Transcription Factors; D004247:DNA