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

Abstract

Summary: 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).

Keywords

• ChIP-seq
• Chromatin immunoprecipitation (ChIP)
• Molecular Biology