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

PLoS One. 2019 Apr 10;14(4):e0215340. doi: 10.1371/journal.pone.0215340. eCollection 2019.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Antibodies*
  • Antibody Specificity
  • Breast Neoplasms / genetics
  • Breast Neoplasms / metabolism
  • Cell Line, Tumor
  • Chromatin Immunoprecipitation Sequencing / methods*
  • Estrogen Receptor alpha / genetics
  • Estrogen Receptor alpha / immunology*
  • Estrogen Receptor alpha / metabolism
  • Female
  • Humans
  • Immunoprecipitation / methods*
  • MCF-7 Cells

Substances

  • Antibodies
  • Estrogen Receptor alpha
  • estrogen receptor alpha, human