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Identification of ChIP-seq and RIME grade antibodies for Estrogen Receptor alpha

Estrogen Receptor alpha (ERα) plays a major role in most breast cancers, and it is the target of endocrine therapies used in the clinic as standard of care for women with breast cancer expressing this receptor. The two methods ChIP-seq (chromatin immunoprecipitation coupled with deep sequencing) and...

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Autores principales: Glont, Silvia-E., Papachristou, Evangelia K., Sawle, Ashley, Holmes, Kelly A., Carroll, Jason S., Siersbaek, Rasmus
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6457525/
https://www.ncbi.nlm.nih.gov/pubmed/30970003
http://dx.doi.org/10.1371/journal.pone.0215340
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author Glont, Silvia-E.
Papachristou, Evangelia K.
Sawle, Ashley
Holmes, Kelly A.
Carroll, Jason S.
Siersbaek, Rasmus
author_facet Glont, Silvia-E.
Papachristou, Evangelia K.
Sawle, Ashley
Holmes, Kelly A.
Carroll, Jason S.
Siersbaek, Rasmus
author_sort Glont, Silvia-E.
collection PubMed
description Estrogen Receptor alpha (ERα) plays a major role in most breast cancers, and it is the target of endocrine therapies used in the clinic as standard of care for women with breast cancer expressing this receptor. The two methods ChIP-seq (chromatin immunoprecipitation coupled with deep sequencing) and RIME (Rapid Immunoprecipitation of Endogenous Proteins) have greatly improved our understanding of ERα function during breast cancer progression and in response to anti-estrogens. A critical component of both ChIP-seq and RIME protocols is the antibody that is used against the bait protein. To date, most of the ChIP-seq and RIME experiments for the study of ERα have been performed using the sc-543 antibody from Santa Cruz Biotechnology. However, this antibody has been discontinued, thereby severely impacting the study of ERα in normal physiology as well as diseases such as breast cancer and ovarian cancer. Here, we compare the sc-543 antibody with other commercially available antibodies, and we show that 06–935 (EMD Millipore) and ab3575 (Abcam) antibodies can successfully replace the sc-543 antibody for ChIP-seq and RIME experiments.
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spelling pubmed-64575252019-05-03 Identification of ChIP-seq and RIME grade antibodies for Estrogen Receptor alpha Glont, Silvia-E. Papachristou, Evangelia K. Sawle, Ashley Holmes, Kelly A. Carroll, Jason S. Siersbaek, Rasmus PLoS One Research Article Estrogen Receptor alpha (ERα) plays a major role in most breast cancers, and it is the target of endocrine therapies used in the clinic as standard of care for women with breast cancer expressing this receptor. The two methods ChIP-seq (chromatin immunoprecipitation coupled with deep sequencing) and RIME (Rapid Immunoprecipitation of Endogenous Proteins) have greatly improved our understanding of ERα function during breast cancer progression and in response to anti-estrogens. A critical component of both ChIP-seq and RIME protocols is the antibody that is used against the bait protein. To date, most of the ChIP-seq and RIME experiments for the study of ERα have been performed using the sc-543 antibody from Santa Cruz Biotechnology. However, this antibody has been discontinued, thereby severely impacting the study of ERα in normal physiology as well as diseases such as breast cancer and ovarian cancer. Here, we compare the sc-543 antibody with other commercially available antibodies, and we show that 06–935 (EMD Millipore) and ab3575 (Abcam) antibodies can successfully replace the sc-543 antibody for ChIP-seq and RIME experiments. Public Library of Science 2019-04-10 /pmc/articles/PMC6457525/ /pubmed/30970003 http://dx.doi.org/10.1371/journal.pone.0215340 Text en © 2019 Glont et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Glont, Silvia-E.
Papachristou, Evangelia K.
Sawle, Ashley
Holmes, Kelly A.
Carroll, Jason S.
Siersbaek, Rasmus
Identification of ChIP-seq and RIME grade antibodies for Estrogen Receptor alpha
title Identification of ChIP-seq and RIME grade antibodies for Estrogen Receptor alpha
title_full Identification of ChIP-seq and RIME grade antibodies for Estrogen Receptor alpha
title_fullStr Identification of ChIP-seq and RIME grade antibodies for Estrogen Receptor alpha
title_full_unstemmed Identification of ChIP-seq and RIME grade antibodies for Estrogen Receptor alpha
title_short Identification of ChIP-seq and RIME grade antibodies for Estrogen Receptor alpha
title_sort identification of chip-seq and rime grade antibodies for estrogen receptor alpha
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6457525/
https://www.ncbi.nlm.nih.gov/pubmed/30970003
http://dx.doi.org/10.1371/journal.pone.0215340
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