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Chromatin immunoprecipitation of transcription factors and histone modifications in Comma-Dbeta mammary epithelial cells

Abstract

Chromatin immunoprecipitation (ChIP) is used to study interactions between proteins and DNA. Nuclear lysates are prepared, and chromatin is fragmented by sonication. Antibodies are used to purify a protein of interest (e.g., a transcription factor or histone mark) along with any bound DNA. The genomic binding sites can then be mapped by sequencing the bound DNA (ChIP-seq) or by qPCR if binding sites are already known. ChIP requires optimization for each cell type, and success is highly antibody dependent. This protocol can be adapted to other cell lines with careful optimization. For complete details on the use and execution of this protocol, please refer to Holliday et al. (2021).

Type Journal
ISBN 2666-1667 (Electronic) 2666-1667 (Linking)
Authors Holliday, H.; Khoury, A.; Swarbrick, A.
Responsible Garvan Author A/Prof Alex Swarbrick
Publisher Name STAR Protocols
Published Date 2021-05-31
Published Volume 2
Published Issue 2
Published Pages 100514
Status Published in-print
DOI 10.1016/j.xpro.2021.100514
URL link to publisher's version https://www.ncbi.nlm.nih.gov/pubmed/34013210