Comprehensive assessment of estrogen receptor beta antibodies in cancer cell line models and tissue reveals critical limitations in reagent specificity

Adam W. Nelson, Arnoud J. Groen, Jodi L. Miller, Anne Y. Warren, Kelly A. Holmes, Gerard A. Tarulli, Wayne D. Tilley, Benita S. Katzenellenbogen, John R Hawse, Vincent J. Gnanapragasam, Jason S. Carroll

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Estrogen Receptor-β (ERβ) has been implicated in many cancers. In prostate and breast cancer its function is controversial, but genetic studies implicate a role in cancer progression. Much of the confusion around ERβ stems from antibodies that are inadequately validated, yet have become standard tools for deciphering its role. Using an ERβ-inducible cell system we assessed commonly utilized ERβ antibodies and show that one of the most commonly used antibodies, NCL-ER-BETA, is non-specific for ERβ. Other antibodies have limited ERβ specificity or are only specific in one experimental modality. ERβ is commonly studied in MCF-7 (breast) and LNCaP (prostate) cancer cell lines, but we found no ERβ expression in either, using validated antibodies and independent mass spectrometry-based approaches. Our findings question conclusions made about ERβ using the NCL-ER-BETA antibody, or LNCaP and MCF-7 cell lines. We describe robust reagents, which detect ERβ across multiple experimental approaches and in clinical samples.

Original languageEnglish (US)
Pages (from-to)138-150
Number of pages13
JournalMolecular and Cellular Endocrinology
Volume440
DOIs
StatePublished - Jan 15 2017

Fingerprint

Estrogen Receptor beta
Estrogen Receptors
Cells
Tissue
Cell Line
Antibodies
Neoplasms
Prostatic Neoplasms
MCF-7 Cells
Mass spectrometry
Mass Spectrometry
Breast

Keywords

  • Antibody
  • Breast
  • Cancer
  • Estrogen receptor beta
  • Prostate

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Endocrinology

Cite this

Comprehensive assessment of estrogen receptor beta antibodies in cancer cell line models and tissue reveals critical limitations in reagent specificity. / Nelson, Adam W.; Groen, Arnoud J.; Miller, Jodi L.; Warren, Anne Y.; Holmes, Kelly A.; Tarulli, Gerard A.; Tilley, Wayne D.; Katzenellenbogen, Benita S.; Hawse, John R; Gnanapragasam, Vincent J.; Carroll, Jason S.

In: Molecular and Cellular Endocrinology, Vol. 440, 15.01.2017, p. 138-150.

Research output: Contribution to journalArticle

Nelson, AW, Groen, AJ, Miller, JL, Warren, AY, Holmes, KA, Tarulli, GA, Tilley, WD, Katzenellenbogen, BS, Hawse, JR, Gnanapragasam, VJ & Carroll, JS 2017, 'Comprehensive assessment of estrogen receptor beta antibodies in cancer cell line models and tissue reveals critical limitations in reagent specificity', Molecular and Cellular Endocrinology, vol. 440, pp. 138-150. https://doi.org/10.1016/j.mce.2016.11.016
Nelson, Adam W. ; Groen, Arnoud J. ; Miller, Jodi L. ; Warren, Anne Y. ; Holmes, Kelly A. ; Tarulli, Gerard A. ; Tilley, Wayne D. ; Katzenellenbogen, Benita S. ; Hawse, John R ; Gnanapragasam, Vincent J. ; Carroll, Jason S. / Comprehensive assessment of estrogen receptor beta antibodies in cancer cell line models and tissue reveals critical limitations in reagent specificity. In: Molecular and Cellular Endocrinology. 2017 ; Vol. 440. pp. 138-150.
@article{996eaab191054392940de8e28f31b36a,
title = "Comprehensive assessment of estrogen receptor beta antibodies in cancer cell line models and tissue reveals critical limitations in reagent specificity",
abstract = "Estrogen Receptor-β (ERβ) has been implicated in many cancers. In prostate and breast cancer its function is controversial, but genetic studies implicate a role in cancer progression. Much of the confusion around ERβ stems from antibodies that are inadequately validated, yet have become standard tools for deciphering its role. Using an ERβ-inducible cell system we assessed commonly utilized ERβ antibodies and show that one of the most commonly used antibodies, NCL-ER-BETA, is non-specific for ERβ. Other antibodies have limited ERβ specificity or are only specific in one experimental modality. ERβ is commonly studied in MCF-7 (breast) and LNCaP (prostate) cancer cell lines, but we found no ERβ expression in either, using validated antibodies and independent mass spectrometry-based approaches. Our findings question conclusions made about ERβ using the NCL-ER-BETA antibody, or LNCaP and MCF-7 cell lines. We describe robust reagents, which detect ERβ across multiple experimental approaches and in clinical samples.",
keywords = "Antibody, Breast, Cancer, Estrogen receptor beta, Prostate",
author = "Nelson, {Adam W.} and Groen, {Arnoud J.} and Miller, {Jodi L.} and Warren, {Anne Y.} and Holmes, {Kelly A.} and Tarulli, {Gerard A.} and Tilley, {Wayne D.} and Katzenellenbogen, {Benita S.} and Hawse, {John R} and Gnanapragasam, {Vincent J.} and Carroll, {Jason S.}",
year = "2017",
month = "1",
day = "15",
doi = "10.1016/j.mce.2016.11.016",
language = "English (US)",
volume = "440",
pages = "138--150",
journal = "Molecular and Cellular Endocrinology",
issn = "0303-7207",
publisher = "Elsevier Ireland Ltd",

}

TY - JOUR

T1 - Comprehensive assessment of estrogen receptor beta antibodies in cancer cell line models and tissue reveals critical limitations in reagent specificity

AU - Nelson, Adam W.

AU - Groen, Arnoud J.

AU - Miller, Jodi L.

AU - Warren, Anne Y.

AU - Holmes, Kelly A.

AU - Tarulli, Gerard A.

AU - Tilley, Wayne D.

AU - Katzenellenbogen, Benita S.

AU - Hawse, John R

AU - Gnanapragasam, Vincent J.

AU - Carroll, Jason S.

PY - 2017/1/15

Y1 - 2017/1/15

N2 - Estrogen Receptor-β (ERβ) has been implicated in many cancers. In prostate and breast cancer its function is controversial, but genetic studies implicate a role in cancer progression. Much of the confusion around ERβ stems from antibodies that are inadequately validated, yet have become standard tools for deciphering its role. Using an ERβ-inducible cell system we assessed commonly utilized ERβ antibodies and show that one of the most commonly used antibodies, NCL-ER-BETA, is non-specific for ERβ. Other antibodies have limited ERβ specificity or are only specific in one experimental modality. ERβ is commonly studied in MCF-7 (breast) and LNCaP (prostate) cancer cell lines, but we found no ERβ expression in either, using validated antibodies and independent mass spectrometry-based approaches. Our findings question conclusions made about ERβ using the NCL-ER-BETA antibody, or LNCaP and MCF-7 cell lines. We describe robust reagents, which detect ERβ across multiple experimental approaches and in clinical samples.

AB - Estrogen Receptor-β (ERβ) has been implicated in many cancers. In prostate and breast cancer its function is controversial, but genetic studies implicate a role in cancer progression. Much of the confusion around ERβ stems from antibodies that are inadequately validated, yet have become standard tools for deciphering its role. Using an ERβ-inducible cell system we assessed commonly utilized ERβ antibodies and show that one of the most commonly used antibodies, NCL-ER-BETA, is non-specific for ERβ. Other antibodies have limited ERβ specificity or are only specific in one experimental modality. ERβ is commonly studied in MCF-7 (breast) and LNCaP (prostate) cancer cell lines, but we found no ERβ expression in either, using validated antibodies and independent mass spectrometry-based approaches. Our findings question conclusions made about ERβ using the NCL-ER-BETA antibody, or LNCaP and MCF-7 cell lines. We describe robust reagents, which detect ERβ across multiple experimental approaches and in clinical samples.

KW - Antibody

KW - Breast

KW - Cancer

KW - Estrogen receptor beta

KW - Prostate

UR - http://www.scopus.com/inward/record.url?scp=85000607279&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85000607279&partnerID=8YFLogxK

U2 - 10.1016/j.mce.2016.11.016

DO - 10.1016/j.mce.2016.11.016

M3 - Article

VL - 440

SP - 138

EP - 150

JO - Molecular and Cellular Endocrinology

JF - Molecular and Cellular Endocrinology

SN - 0303-7207

ER -