Proteomics,Multiomics

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

ABSTRACT: Estrogen Receptor alpha (ERα) is a key driver of most breast cancers, and it is the target of endocrine therapies used in the clinic to treat women with ERα positive (ER+) breast cancer. 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 to pull down 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.

OTHER RELATED OMICS DATASETS IN: GSE128208

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Evangelia Papachristou  

LAB HEAD: Jason Carroll

PROVIDER: PXD012930 | Pride | 2019-03-21

REPOSITORIES: Pride

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

Glont Silvia-E SE   Papachristou Evangelia K EK   Sawle Ashley A   Holmes Kelly A KA   Carroll Jason S JS   Siersbaek Rasmus R  

PloS one 20190410 4

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 componen  ...[more]

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