Methods for Detection of NOX-Derived Superoxide Radical Anion and Hydrogen Peroxide in Cells

NADPH oxidases (NOX) are transmembrane enzymes, which catalyze the formation of reactive oxygen species (ROS). In humans and most mammals, the NOX family comprises seven members, namely, NOX1-5 and the dual oxidases DUOX1 and 2. The primary product of most NOX isoforms is the superoxide radical anio...

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Published in:Methods in molecular biology (Clifton, N.J.) Vol. 1982; p. 233
Main Authors: Augsburger, Fiona, Filippova, Aleksandra, Jaquet, Vincent
Format: Journal Article
Language:English
Published: United States 2019
Subjects:
Animals
Anions - metabolism
CHO Cells
Cricetulus
Enzyme Assays
HEK293 Cells
Humans
Hydrogen Peroxide - metabolism
Loss of Function Mutation
NADPH Oxidases - genetics
NADPH Oxidases - metabolism
Reactive Oxygen Species - metabolism
Superoxides - metabolism
Hydrogen peroxide
Superoxide radical anion
NADPH oxidase
Cellular assay
Peroxidase
WST-1
Amplex red
ISSN:1940-6029, 1940-6029
Online Access:Get more information
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Summary:NADPH oxidases (NOX) are transmembrane enzymes, which catalyze the formation of reactive oxygen species (ROS). In humans and most mammals, the NOX family comprises seven members, namely, NOX1-5 and the dual oxidases DUOX1 and 2. The primary product of most NOX isoforms is the superoxide radical anion O , which is rapidly dismutated in hydrogen peroxide (H O ), while NOX4 and DUOX mostly generate H O ROS are multifunctional molecules in tissues, and NOX-derived ROS cellular functions are as diverse as microbial killing (NOX2), thyroid hormone synthesis (DUOX2), or otoconia formation in the inner ear (NOX3). NOX are potential pharmacological targets in numerous diseases such as diabetes, fibrosis, and brain ischemia, and NOX inhibitors are currently under development. Here we describe two cellular assays to detect extracellular O and H O in cells overexpressing specific NOX isoforms and their subunits.
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ISSN:1940-6029
1940-6029
DOI:10.1007/978-1-4939-9424-3_13