Acquisition of temporal HIF transcriptional activity using a secreted luciferase assay

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

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Pages37-44
Number of pages8
Volume1742
DOIs
Publication statusPublished - 13 Jan 2018

Publication series

NameMethods in Molecular Biology
Volume1742
ISSN (Print)1064-3745

Keywords

  • Activity
  • HIF
  • Hypoxia
  • Luciferase
  • Time-resolved
  • Transcription

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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 Methods in Molecular Biology (Vol. 1742, pp. 37-44). (Methods in Molecular Biology; Vol. 1742). Humana Press. https://doi.org/10.1007/978-1-4939-7665-2_4