Superoxide dismutase (SOD) is an antioxidant enzyme involved in defense against reactive oxygen species (ROS). SOD catalyzes the dismutation of superoxide radical anion (O2-) to hydrogen peroxide, which is then catalyzed to O2 and H2O by glutathione peroxidase and/or catalase. Several classes of SOD have been discovered and can be differentiated according to the metal associated with their active site as well as their location within a cell or tissue. In mammals, Cu/ZnSOD (SOD1) is localized in the cytosol and ecSOD (SOD3) in the extracellular matrix of various tissues. MnSOD or SOD2 exists as a tetramer with individual subunit MW of approximately 23 kDa and Mn at its active site. It is initially synthesized with a leader peptide which targets this enzyme exclusively to the mitochondrial spaces. MnSOD, as the primary antioxidant enzyme for scavenging superoxide radicals in mitochondria, is essential for the survival of all aerobic organisms. Dysregulation of MnSOD in the form of under-expression is being investigated relative to various disease states as well as aging. Overexpression of MnSOD has been shown to protect against oxidative stress induced cell death and tissue injury. MnSOD has also been shown to play a major role in promoting cellular differentiation and tumorigenesis and in protecting against hyperoxia-induced pulmonary toxicity.
The NWLSS™ MnSOD Assay is based on a sandwich Enzyme-Linked Immunosorbent Assay (ELISA). The microtiter plate provided in this kit has been pre-coated with a monoclonal antibody specific to human MnSOD. This stationary phase antibody binds sample or standard MnSOD while nonbound proteins are removed by washing. Next, bound MnSOD is tagged with a biotin-conjugated monoclonal antibody specific for MnSOD followed by Avidin conjugated to Horseradish Peroxidase (HRP). Subsequent addition of TMB-substrate solution causes blue color (650 nm) development proportional to the amount of MnSOD originally captured by the stationary phase antibody. Finally, addition of a sulfuric acid solution stops the reaction resulting in a yellow color product measured at 450 nm. Sample MnSOD concentration is determined by comparing the 450 nm absorbance of sample wells to the absorbance of known standards.
||96 Well Sandwich ELISA (6 X 16 well strips in frame)
||Human cell lysate or tissue homogenate
||Precoated Microwell Strips
rHu MnSOD Standard
Sample/Standard Dilution Buffer
Reagent Dilution Buffer
Assay Preparation Buffer
Biotin Labeled anti hu MnSOD