A Novel Dual Antibody Staining Assay to Measure Estrogen Receptor Transcriptional Activity.

Journal of Fluorescence
F. van HemertAnja van de Stolpe

Abstract

Activity of the canonical estrogen receptor (ER) pathway is equivalent to functional activity of the nuclear ER transcription factor. Monoclonal antibodies (MoAbs) that identify nuclear ER in cells and tissue samples are frequently used to assess ER transcriptional activity, however, it remains unclear if this approach is sufficiently predictive of ER pathway activity. This study uses ER-positive breast cancer cell lines (MCF7 and T47D) in which ER transcriptional activity was quantified using an mRNA-based ER pathway activity assay. The relationship between ER activity and nuclear ER staining with ER MoAbs was then investigated. Confirming earlier findings, the results show that while the presence of ER in the cell nucleus is a prerequisite for ER activity, it is not predictive of ER transcriptional activity. There were remarkable differences in the behaviours of the antibodies used in the study. EP1 and 1D5 showed reduced nuclear staining when ER was transcriptionally active, while staining with H4624 was independent of ER activity. To improve discrimination between active and inactive nuclear ER based on ER staining, a method was developed which consists of dual ER MoAb immunofluorescent staining, followed by generation of a...Continue Reading

References

Dec 1, 1993·The Journal of Steroid Biochemistry and Molecular Biology·B S KatzenellenbogenC K Wrenn
Nov 1, 2016·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Carla Rossana ScaliaGiorgio Cattoretti

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Methods Mentioned

BETA
BioDetection
Fluorescence

Software Mentioned

Seaborn
matplotlib
Pandas
learn
MATLAB
scikit

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