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Acquisition of Temporal HIF Transcriptional Activity Using a Secreted Luciferase Assay

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Hypoxia

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1742))

Abstract

Here we describe a simple method based on secreted luciferase driven by a hypoxia-inducible factor (HIF) response element (HRE) that allows the acquisition of dynamic and high-throughput data on HIF transcriptional activity during hypoxia and pharmacological activation of HIF. The sensitivity of the assay allows for the secreted luciferase to be consecutively sampled (as little as 1% of the total supernatant) over an extended time period, thus allowing the acquisition of time-resolved HIF transcriptional activity.

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Acknowledgments

This work was supported by Science Foundation Ireland (grant number 06/CE/B1129).

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Authors and Affiliations

  1. Systems Biology Ireland, University College Dublin, Dublin, Ireland

    Miguel A. S. Cavadas, Cormac T. Taylor & Alex Cheong

  2. Conway Institute of Biomolecular and Biomedical Research, School of Medicine, University College Dublin, Dublin, Ireland

    Miguel A. S. Cavadas, Cormac T. Taylor & Alex Cheong

  3. Instituto Gulbenkian de Ciencia, Oeiras, Portugal

    Miguel A. S. Cavadas

  4. Life and Health Sciences, Aston University, Birmingham, UK

    Alex Cheong

Authors
  1. Miguel A. S. Cavadas
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  2. Cormac T. Taylor
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  3. Alex Cheong
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Corresponding author

Correspondence to Alex Cheong .

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Editors and Affiliations

  1. Department of Neurosurgery, Clinical Neurosciences Center, University of Utah, Salt Lake City, Utah, USA

    L. Eric Huang

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Cavadas, M.A.S., Taylor, C.T., Cheong, A. (2018). Acquisition of Temporal HIF Transcriptional Activity Using a Secreted Luciferase Assay. In: Huang, L. (eds) Hypoxia. Methods in Molecular Biology, vol 1742. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7665-2_4

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  • DOI: https://doi.org/10.1007/978-1-4939-7665-2_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7664-5

  • Online ISBN: 978-1-4939-7665-2

  • eBook Packages: Springer Protocols

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