title: Cell-Based Assay for Identifying the Modulators of Antioxidant Response Element Signaling Pathway.
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.
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Building C, MSC: 3375, 9800 Medical Center Drive, Bethesda, MD, 20892, USA.
authors: Jinghua
authors: J
authors: Zhao
authors: Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA.
authors: Sunita J
authors: SJ
authors: Shukla
authors: 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.
authors: Menghang
authors: M
authors: Xia
mesh_terms: D061745:Antioxidant Response Elements; D000975:Antioxidants / Q000494:pharmacology*; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D056945:Hep G2 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000235:genetics / Q000378:metabolism; D051267:NF-E2-Related Factor 2 / Q000235:genetics* / Q000378:metabolism; D016877:Oxidants / Q000494:pharmacology*; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000187:drug effects; D011401:Promoter Regions, Genetic; D017382:Reactive Oxygen Species / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015398:Signal Transduction; D001618:beta-Lactamases / Q000235:genetics / Q000378:metabolism; D019324:beta-Naphthoflavone / Q000494:pharmacology
chemical_list: D000975:Antioxidants; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D016877:Oxidants; D017382:Reactive Oxygen Species; D019324:beta-Naphthoflavone; D016660:NAD(P)H Dehydrogenase (Quinone); C447479:NQO1 protein, human; D001618:beta-Lactamases
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)