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Methods Mol Biol. 2016;1473:55-62. doi: 10.1007/978-1-4939-6346-1_6.

Cell-Based Assay for Identifying the Modulators of Antioxidant Response Element Signaling Pathway.

Author information

1
National Center for Advancing Translational Sciences, National Institutes of Health, Building C, MSC: 3375, 9800 Medical Center Drive, Bethesda, MD, 20892, USA.
2
Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA.
3
National Center for Advancing Translational Sciences, National Institutes of Health, Building C, MSC: 3375, 9800 Medical Center Drive, Bethesda, MD, 20892, USA. mxia@mail.nih.gov.

Abstract

The antioxidant response element (ARE) signaling pathway plays an important role in the amelioration of cellular oxidative stress. Thus, assays that detect this pathway can be useful for identifying chemicals that induce or inhibit oxidative stress signaling. The focus of this chapter is to describe a cell-based ARE assay in a quantitative high-throughput screening (qHTS) format to test a large collection of compounds that induce nuclear factor erythroid 2-related factor (Nrf2)/ARE signaling. The assay is described through cell handling, assay preparation, and instrument usage.

KEYWORDS:

Antioxidant response element (ARE); Fluorescence Resonance Energy Transfer (FRET); Nuclear factor erythroid 2-related factor (Nrf2); Quantitative high-throughput screening (qHTS); Reactive oxygen species (ROS); β-lactamase (bla)

PMID:
27518623
DOI:
10.1007/978-1-4939-6346-1_6
[Indexed for MEDLINE]

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