A rapid micro chromatin immunoprecipitation assay (microChIP).
Interactions of proteins with DNA mediate many critical nuclear functions. Chromatin immunoprecipitation (ChIP) is a robust technique for studying protein-DNA interactions. Current ChIP assays, however, either require large cell numbers, which prevent their application to rare cell samples or small-tissue biopsies, or involve lengthy procedures. We describe here a 1-day micro ChIP (microChIP) protocol suitable for up to eight parallel histone and/or transcription factor immunoprecipitations from a single batch of 1,000 cells. MicroChIP technique is also suitable for monitoring the association of one protein with multiple genomic sites in 100 cells. Alterations in cross-linking and chromatin preparation steps also make microChIP applicable to approximately 1-mm(3) fresh- or frozen-tissue biopsies. From cell fixation to PCR-ready DNA, the procedure takes approximately 8 h for 16 ChIPs.
Dahl
John Arne
JA
Department of Biochemistry, Faculty of Medicine, Institute of Basic Medical Sciences, University of Oslo, Oslo 0317, Norway.
Collas
Philippe
P
D000818:Animals; D000906:Antibodies; D002452:Cell Count; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D003432:Cross-Linking Reagents; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D006657:Histones / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D008832:Microchemistry / Q000379:methods; D016133:Polymerase Chain Reaction; D012319:RNA Polymerase II / Q000276:immunology / Q000378:metabolism; D013997:Time Factors
D000906:Antibodies; D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D006657:Histones; D004247:DNA; D012319:RNA Polymerase II