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Use of the monoclonal antibody DAKO-ERβ (8D5-1) to measure oestrogen receptor beta in breast cancer cells

Lookup NU author(s): Fiona Girdler, Dr Brian Shenton, Dr James Young, Dr Ian Brotherick


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Background: Oestrogen receptor beta (ERβ) is highly homologous with the classical ER (known now as ERα). The exact role of ERβ in breast cancer and its contribution in influencing patient response to endocrine therapy remains unclear. The aim of this study was to develop and evaluate a flow cytometric method for the detection of ERβ in breast cancer cells using the DAKO monoclonal anti-ERβ 8D5-1 antibody. Methods: MCF7 cells were used as a positive control and U937 as a negative control for titration of the antibody. Cell lines and tumour samples were fixed with 1% paraformaldehyde and permeabilised with 0.5% saponin prior to flow cytometric analysis. Results: A ten fold difference in expression of ERβ within the different breast cell lines studied was found. Confirmation of antibody specificity against ERβ protein by Western blot analysis detected a single band at approximately 65kDa. ERβ immunopositive nuclei were identified in MCF7 cells by immunohistochemistry. Conclusions: DAKO ERβ 8D5-1 antibody is specific for ERβ protein and does not cross react with ERα protein. Using this antibody, ERβ can be detected and accurately quantified in cell lines and solid breast tumours by flow cytometry. © 2001 Wiley-Liss, Inc.

Publication metadata

Author(s): Girdler F, Browell DA, Cunliffe WJ, Shenton BK, Hemming JD, Scorer P, Young JR, Brotherick I

Publication type: Article

Publication status: Published

Journal: Cytometry

Year: 2001

Volume: 45

Issue: 1

Pages: 65-72

ISSN (print): 1552-4922

ISSN (electronic): 1552-4930

Publisher: John Wiley & Sons


DOI: 10.1002/1097-0320(20010901)45:1<65::AID-CYTO1145>3.0.CO;2-8

PubMed id: 11598948


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