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

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Nelson, AW; Groen, AJ; Miller, JL; Warren, AY; Holmes, KA; Tarulli, GA; Tilley, WD; Katzenellenbogen, BS; Hawse, JR; Gnanapragasam, VJ; ...Date
2017-01-15Source Title
Molecular and Cellular EndocrinologyPublisher
ELSEVIER IRELAND LTDUniversity of Melbourne Author/s
Tarulli, GerardAffiliation
School of BioSciencesMetadata
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Nelson, A. W., Groen, A. J., Miller, J. L., Warren, A. Y., Holmes, K. A., Tarulli, G. A., Tilley, W. D., Katzenellenbogen, B. S., Hawse, J. R., Gnanapragasam, V. J. & Carroll, J. S. (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, 440 (C), pp.138-150. https://doi.org/10.1016/j.mce.2016.11.016.Access Status
Open AccessAbstract
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.
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