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Superoxide Measurement

HG Huiming Gao

Published: Nov 20, 2011 DOI: 10.21769/BioProtoc.155 Views: 13556

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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.

Materials and Reagents

  1. Phenol red-free HBSS (Life Technologies, InvitrogenTM, catalog number: 14175-095 )
  2. Superoxide dismutase from bovine erythrocytes (Sigma-Aldrich, catalog number: S7571-30KU )
  3. Phorbol 12-myristate 13-acetate (PMA)
  4. Lipopolysaccharide (LPS)
  5. Fresh WST-1 solution (see Recipes)
  6. SOD stock solution (see Recipes)

Equipment

  1. Microplate spectrophotometer (SpectraMax Plus)
  2. 96-well plates

Procedure

Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.

Category

Neuroscience > Cellular mechanisms

Biochemistry > Other compound > Reactive oxygen species

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1 Q&A

recipe for WST-1 PREPARATION PLEASE
edit 0 Answer 0 View Apr 13, 2018