Protocol abstract
Nature Protocols 1, - 179 - 185 (2006)
Published online: 27 June 2006 | doi:10.1038/nprot.2006.27
Subject Categories: Biochemistry and protein analysis | Immunological techniques | Isolation, purification and separation | Nucleic acid based molecular biology
Protocol for the fast chromatin immunoprecipitation (ChIP) method
Joel D Nelson1,2, Oleg Denisenko2 & Karol Bomsztyk1,2
Abstract
Chromatin and transcriptional processes are among the most intensively studied fields of biology today. The introduction of chromatin immunoprecipitations (ChIP) represents a major advancement in this area. This powerful method allows researchers to probe specific protein-DNA interactions in vivo and to estimate the density of proteins at specific sites genome-wide. We have introduced several improvements to the traditional ChIP assay, which simplify the procedure, greatly reducing the time and labor required to complete the assay. The simplicity of the method yields highly reproducible results. Our improvements facilitate the probing of multiple proteins in a single experiment, which allows for the simultaneous monitoring of many genomic events. This method is particularly useful in kinetic studies where multiple samples are processed at the same time. Starting with sheared chromatin, PCR-ready DNA can be isolated from 16–24 ChIP samples in 4–6 h using the fast method.
- Molecular and Cellular Biology Program, University of Washington, Seattle, Washington 98109, USA.
- UW Medicine at Lake Union, University of Washington, Seattle, Washington 98109, USA.
Correspondence to: Karol Bomsztyk1,2 e-mail: karolb@u.washington.edu
nature-products
A-Z product listing
- anti–acetylated histone H3 (anti–H3-K9Ac(Cell Signaling)
- anti–CREB binding protein (anti-CBP(Santa Cruz Biotechnology)
- anti–dimethyl H3 (anti–H3-K4m2(Upstate Biotechnology)
- anti–H3 lysine 27 trimethylation (anti–H3-K27m3(Abcam)
- Anti–histone H3(Abcam)
- Anti–RNA polymerase II(Santa Cruz Biotechnology)
