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

被引:8
|
作者
Glont, Silvia-E. [1 ]
Papachristou, Evangelia K. [1 ]
Sawle, Ashley [1 ]
Holmes, Kelly A. [1 ]
Carroll, Jason S. [1 ]
Siersbaek, Rasmus [1 ]
机构
[1] Univ Cambridge, Canc Res UK Cambridge Inst, Robinson Way, Cambridge, England
来源
PLOS ONE | 2019年 / 14卷 / 04期
关键词
GENOME-WIDE ANALYSIS; BREAST-CANCER; BINDING; FOXA1;
D O I
10.1371/journal.pone.0215340
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Estrogen Receptor alpha (ERa) 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 ERa 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 ERa have been performed using the sc-543 antibody from Santa Cruz Biotechnology. However, this antibody has been discontinued, thereby severely impacting the study of ERa 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.
引用
收藏
页数:10
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