Protocol abstract
Nature Protocols 3, - 1032 - 1045 (2008)
Published online: 29 May 2008 | doi:10.1038/nprot.2008.68
Subject Categories: Genetic analysis | Immunological techniques | Isolation, purification and separation | Nucleic acid based molecular biology
A rapid micro chromatin immunoprecipitation assay (ChIP)
John Arne Dahl1 & Philippe Collas1
Abstract
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 (
ChIP) protocol suitable for up to eight parallel histone and/or transcription factor immunoprecipitations from a single batch of 1,000 cells. ChIP 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 ChIP applicable to 
1-mm3 fresh- or frozen-tissue biopsies. From cell fixation to PCR-ready DNA, the procedure takes 8 h for 16 ChIPs.
- Department of Biochemistry, Faculty of Medicine, Institute of Basic Medical Sciences, University of Oslo, Oslo 0317, Norway.
Correspondence to: Philippe Collas1 e-mail: philippe.collas@medisin.uio.no
nature-products
A-Z product listing
- 0.6-ml Centrifuge tubes(Axygen)
- 1 M Tris–HCl, pH 7.5(Sigma-Aldrich)
- 1 M Tris–HCl, pH 8.0(Sigma-Aldrich)
- 200-l PCR tubes in 8-tube strip format(Axygen)
- 25:24:1(Invitrogen)
- 3 M NaAc(Sigma-Aldrich)
