Evaluation of seven oestrogen receptor β antibodies for immunohistochemistry, western blotting, and flow cytometry in human breast tissue

G. P. Skliris, A. T. Parkes, J. L. Limer, S. E. Burdall, P. J. Carder, V. Speirs*

*Corresponding author for this work

Research output: Contribution to journalArticle

113 Citations (Scopus)

Abstract

Two oestrogen receptors, ERα and ERβ, exist. While much is known about ERα, the role of ERβ is still undefined, especially at the protein level. The aim of this study was to determine the utility of seven ERβ antibodies (14C8, 8D5, PAI313, PPG5/10, N19, 9.88, and D7N) raised against different domains of ERβ in three commonly used laboratory applications, namely immunohistochemistry, western blot, and flow cytometry, using human breast material. For immunohistochemical analysis of frozen material, PAI313 and D7N gave stronger and more specific signals than 14C8, 8D5, and PPG5/10. In paraffin sections, 14C8, closely followed by PPG5/10, gave by far the most superior nuclear immunoreactivity, compared with the other antibodies tested. In general, flow cytometry results mirrored the immunohistochemistry data for paraffin sections, with antibodies ranked 14C8>8D5 ≥ PAI-313 > PPG5/10>D7N. For western blotting, 8D5 and D7N yielded the strongest and most consistent bands, with weaker bands seen with the others. It is concluded that ERβ protein can be detected using specific antibodies. However, there is considerable variation between the specificity and application of these antibodies, highlighting the fact that careful optimization is required when selecting an antibody for use in a particular laboratory technique.

Original languageEnglish
Pages (from-to)155-162
Number of pages8
JournalJournal of Pathology
Volume197
Issue number2
DOIs
Publication statusPublished - 2002

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Estrogen Receptors
Flow Cytometry
Breast
Western Blotting
Immunohistochemistry
Antibodies
Paraffin
Antibody Specificity
Proteins

Keywords

  • ERβ
  • Flow cytometry
  • Immunoblotting
  • Immunohistochemistry
  • Mammary gland

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Evaluation of seven oestrogen receptor β antibodies for immunohistochemistry, western blotting, and flow cytometry in human breast tissue. / Skliris, G. P.; Parkes, A. T.; Limer, J. L.; Burdall, S. E.; Carder, P. J.; Speirs, V.

In: Journal of Pathology, Vol. 197, No. 2, 2002, p. 155-162.

Research output: Contribution to journalArticle

Skliris, G. P. ; Parkes, A. T. ; Limer, J. L. ; Burdall, S. E. ; Carder, P. J. ; Speirs, V. / Evaluation of seven oestrogen receptor β antibodies for immunohistochemistry, western blotting, and flow cytometry in human breast tissue. In: Journal of Pathology. 2002 ; Vol. 197, No. 2. pp. 155-162.
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