Hunter, AL;
Narang, N;
Baxter, M;
Ray, DW;
Poolman, TM;
(2019)
An improved method for quantitative ChIP studies of nuclear receptor function.
Journal of Molecular Endocrinology
, 62
(4)
pp. 169-177.
10.1530/JME-18-0243.
Preview |
Text
[14796813 - Journal of Molecular Endocrinology] An improved method for quantitative ChIP studies of nuclear receptor function.pdf - Published Version Download (1MB) | Preview |
Abstract
Chromatin immunoprecipitation (ChIP) is a valuable tool for the endocrine researcher, providing a means to measure the 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.
| Type: | Article |
|---|---|
| Title: | An improved method for quantitative ChIP studies of nuclear receptor function |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1530/JME-18-0243 |
| Publisher version: | https://doi.org/10.1530/JME-18-0243 |
| Language: | English |
| Additional information: | Copyright © The authors 2019. This work is licensed under a Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/). |
| Keywords: | ChIP; digital PCR; ddPCR; troubleshooting; nuclear receptors |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Structural and Molecular Biology |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10103184 |
Archive Staff Only
 |
View Item |
