An improved method for quantitative ChIP studies of nuclear receptor function

Research output: Contribution to journalArticle

Abstract

Chromatin immunoprecipitation (ChIP) is a valuable tool for the endocrine researcher, providing a means to measure recruitment of hormone-activated nuclear receptors, for example. However, the technique can be challenging to perform and has multiple experimental steps, risking introduction of error at each. The data produced can be challenging to interpret; several different methods are commonly used for normalising data and thus comparing between conditions. Absolute, sensitive quantification of protein-bound DNA is important for correct interpretation of the data. In addition, such quantification can help the investigator in troubleshooting experiments. Here, we outline a ChIP strategy combining droplet digital PCR for accurate quantification with an internal spike-in control for normalisation. This combination strengthens the reliability of ChIP data and allows the operator to optimise their protocol with greater confidence

Bibliographical metadata

Original languageEnglish
Pages (from-to)169-177
Number of pages8
JournalJ Mol Endocrinol
Volume62
Issue number4
Early online date27 Mar 2019
DOIs
Publication statusPublished - 1 May 2019