The NWLSS™ Superoxide Oxidant Status Assay is recommended for the determination of superoxide in isolated monocyte populations and general oxidant levels in other biological samples.
Superoxide and other reactive oxygen species play very important role in diseases and pathology related to oxidative stress. Their extremely low steady-state concentration in-vivo require sensitive methods of measurement. A number of techniques have been described in the literature including ferricytochrome c reduction, nitroblue tetrazolium reduction, aconitase activity inhibition, nitrone spin trapping, electrochemical detection, and chemiluminescent assays with luminol and lucigenin. The luminol based assay has been criticized as a specific quantitative method to determine superoxide production in cells due to its sensitivity to a number of reactive oxygen species (hypochlorite, peroxynitrite and hydroxyl radical from H2O2+metal/heme-proteins). Also, the first intermediate in the reaction can react with oxygen to generate superoxide resulting in higher apparent superoxide production rate than actual cellular production rate. However, this assay can provide a valid, simple and sensitive gross assessment of reactive oxygen species in many biological sample types and is an important method in the analysis of specific superoxide production by monocytes.
The NWLSS™ NWK-SOS01 method is based on Luminol reaction with superoxide to produce a luminophore with an emission peak at ~425 nm. Orthovanadate is used to increase luminescence respose by uo to 50X. The luminescence intensity is proportional to the amount of superoxide in the sample. The figure below shows the principle of the reaction.
View Assay Protocol
A chemiluminescent method for determination superoxide levels in monocyte cells and provides data on general oxidant levels in other biological samples
Isolated cell populations, whole blood, tissue homogenates
Superoxide when measuring isolated monocyte populations else general oxidant levels in biological samples.
LLD = 0.3 U/mL in Reaction Mix|
5 U/mL in Sample Added to Reaction Mix
|Storage and Stability:||
Six months from date of manufacture when stored at 2-8C
HBSS Buffer Solution
Phorbol Myristate Acetate (PMA)
PMA Solvent (DMSO)