Abstract
In situ proximity ligation assay (PLA) is a powerful method for detection, localization, and quantification of proteins, protein–protein interactions, and posttranslational modifications. Proteins detected by two specific antibodies are recognized by proximity probes conjugated with complementary oligonucleotides to allow the formation of circular DNA probes when bound in close proximity. Subsequent amplification of this DNA can then be visualized. Here, we describe the in situ PLA method for the detection of the ERα/Src/PI3K complex in breast cancer. We used two different techniques for detecting the signals: fluorescent detection for cell line analysis and bright-field revelation, which is better suited to clinical analysis of patient samples.
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Acknowledgments
This study was partially supported by the “Fondation Arc Cancer” (ARC) and Fondation de France (FDF). C.P. was supported by the French Ministry of Research and by ARC. J.R. was supported by FDF.
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Poulard, C., Rambaud, J., Le Romancer, M., Corbo, L. (2014). Proximity Ligation Assay to Detect and Localize the Interactions of ERα with PI3-K and Src in Breast Cancer Cells and Tumor Samples. In: Castoria, G., Auricchio, F. (eds) Steroid Receptors. Methods in Molecular Biology, vol 1204. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-1346-6_12
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DOI: https://doi.org/10.1007/978-1-4939-1346-6_12
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-1345-9
Online ISBN: 978-1-4939-1346-6
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