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AlphaScreen-Based Assays: Ultra-High-Throughput Screening for Small-Molecule Inhibitors of Challenging Enzymes and Protein-Protein Interactions

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High Throughput Screening

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1439))

Abstract

AlphaScreen technology has been routinely utilized in high-throughput screening assays to quantify analyte accumulation or depletion, bimolecular interactions, and post-translational modifications. The high signal-to-background, dynamic range, and sensitivity associated with AlphaScreens as well as the homogenous assay format and reagent stability make the technology particularly well suited for high-throughput screening applications. Here, we describe the development of AlphaScreen assays to identify small-molecule inhibitors of enzymes and protein-protein interactions using the highly miniaturized 1536-well format. The subsequent implementation of counter assays to identify false-positive compounds is also discussed.

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Acknowledgement

This work was supported by the NCATS Division of Pre-Clinical Innovation Intramural Program.

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Authors and Affiliations

  1. National Center for Advancing Translational Sciences, National Institutes of Health, 9800 Medical Center Drive, Building B, Rm. 1034E, Rockville, MD, 20850, USA

    Adam Yasgar, Ajit Jadhav, Anton Simeonov & Nathan P. Coussens

Authors
  1. Adam Yasgar
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  2. Ajit Jadhav
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  3. Anton Simeonov
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  4. Nathan P. Coussens
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Corresponding author

Correspondence to Nathan P. Coussens .

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Editors and Affiliations

  1. Epizyme, Inc., Cambridge, Massachusetts, USA

    William P. Janzen

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© 2016 Springer Science+Business Media New York

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Yasgar, A., Jadhav, A., Simeonov, A., Coussens, N.P. (2016). AlphaScreen-Based Assays: Ultra-High-Throughput Screening for Small-Molecule Inhibitors of Challenging Enzymes and Protein-Protein Interactions. In: Janzen, W. (eds) High Throughput Screening. Methods in Molecular Biology, vol 1439. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3673-1_5

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  • DOI: https://doi.org/10.1007/978-1-4939-3673-1_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3671-7

  • Online ISBN: 978-1-4939-3673-1

  • eBook Packages: Springer Protocols

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