An optimized two-step chromatin immunoprecipitation protocol to quantify the associations of two separate proteins and their common target DNA

Lingli He; Wentao Yu; Wenxiang Zhang; Lei Zhang

doi:10.1016/j.xpro.2021.100504

An optimized two-step chromatin immunoprecipitation protocol to quantify the associations of two separate proteins and their common target DNA

STAR Protoc. 2021 Apr 27;2(2):100504. doi: 10.1016/j.xpro.2021.100504. eCollection 2021 Jun 18.

Authors

Lingli He¹, Wentao Yu¹, Wenxiang Zhang¹, Lei Zhang^{1

2

3}

Affiliations

¹ State Key Laboratory of Cell Biology, Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China.
² School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China.
³ School of Life Science, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou 310024, China.

Abstract

Sequential chromatin immunoprecipitation (ChIP) is commonly used to investigate DNA-protein and protein-protein interactions to a specific genomic region. However, it can be tricky to achieve a robust and reproducible signal with sequential ChIP. Here, we provide an optimized two-step ChIP protocol to quantify the in vivo associates of multiple proteins with the same DNA regulatory element. For complete details on the use and execution of this protocol, please refer to He et al. (2020).

Keywords: Chromatin immunoprecipitation (ChIP); Molecular Biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line
Chromatin Immunoprecipitation*
DNA / chemistry
DNA / metabolism*
DNA-Binding Proteins / chemistry
DNA-Binding Proteins / metabolism*
Humans
Response Elements*

Substances

DNA-Binding Proteins
DNA