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Superoxide Measurement
Published: Nov 20, 2011 DOI: 10.21769/BioProtoc.155 Views: 13568
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Abstract
Superoxide dismutase (SOD)-inhibitable reduction of tetrazolium salt, WST-1, is used to measure extracellular superoxide free radical release from various immune cells such as macrophage, neutrophils, microglia (brain macrophage) after stimulation. This protocol has been developed and improved over the years by various researchers in Dr. Hong’s lab, especially Dr. Bin Liu.
Background
Materials and Reagents
- Phenol red-free HBSS (Life Technologies, InvitrogenTM, catalog number: 14175-095 )
- Superoxide dismutase from bovine erythrocytes (Sigma-Aldrich, catalog number: S7571-30KU )
- Phorbol 12-myristate 13-acetate (PMA)
- Lipopolysaccharide (LPS)
- Fresh WST-1 solution (see Recipes)
- SOD stock solution (see Recipes)
Equipment
- Microplate spectrophotometer (SpectraMax Plus)
- 96-well plates
Procedure
Category
Neuroscience > Cellular mechanisms
Biochemistry > Other compound > Reactive oxygen species
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