Superoxide dismutase (SOD) is an antioxidant enzyme involved in the defense system against reactive oxygen species (ROS). SOD catalyzes the dismutation reaction of superoxide radical anion (O -) to hydrogen peroxide, which is then catalyzed to innocuous O and H O by 222 glutathione peroxidase and catalase. Several classes of SOD have been identified. : Cu/Zn SOD (SOD1) is localized in cytosol, Mn SOD (SOD2) in mitochondria, and ecSOD (SOD3) in extracellular space. Cu/Zn SOD (SOD1) is a homodimer containing copper and zinc that is found almost exclusively in intracellular cytoplasmic spaces. The level of SOD1 elevates in response to a wide array of mechanical, chemical and biological messengers such as heat shock, shear stress, and UVB- and X-irradiation. Decreased levels of SOD1 expression can also be triggered by activation of the AP2 transcription factor. A down-regulation of SOD1 has been shown in alveolar type II epithelial cells and lung fibroblasts after exposure to hypoxia. The genomic organization of SOD1 genes shows striking similarities between species and consists of five exons and four introns. The TATA and CCAAT boxes, as well as several highly conserved GC-rich regions, have been localized in the proximal promoter region. Also, more than 90 different mutations of the SOD1 gene have been associated with amyotrophic lateral sclerosis (ALS), also known as Lou Gehrig's disease.Intended Use:Human Superoxide Dismutase-1 (human SOD1) ELISA kit is to be used for the in vitro quantitative determination of human SOD1 in human serum, human plasma, cell lysate and buffered solution. The assay will recognize both native and recombinant human SOD1.This kit has been configured for research use only and is not to be used in diagnostic procedures.Range:12.5-800pg/mlSensitivity:The minimal detectable dose of human SOD1 was calculated to be 12.5pg/ml, by subtracting two standard deviations from the mean of 10 zero standard replicates (ELISA buffer, S0) and intersecting this value with the standard curve obtained in the same calculation.Specificity:The following substances were tested and found to have no cross-reactivity: human SOD2, SOD3, SOD4, rat SOD1 and mouse SOD1.Test Principle:The design of this 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 SOD1. Samples are pippetted into these wells. Nonbound SOD1 and other components of the sample should be removed by washing, then biotin-conjugated monoclonal antibody specific to SOD1 added. In order to quantitatively determine the amount of SOD1 present in the sample, Avidin conjugated to Horseradish Peroxidase (HRP) should be added to each microplate well. The final step, a TMB-substrate solution added to each well. Finally, a sulfuric acid solution is added and the resulting yellow colored product is measured at 450nm. Since the increases in absorbency is directly proportional to the amount of captured SOD1.Kit Components:S8060-32P1: Microtiter plate, 1x96 wells *S8060-32P2: Standard, recombinant human SOD1 (lyophilized), 1x1vial S8060-32P3: Secondary Antibody (Biotin), 100x (lyophilized), 1x1vialS8060-32P4: Avidin (HRP), 100x, 1x150ulS8060-32P5: Incubation Buffer, 1x30mlS8060-32P6: Wash Buffer, 20x, 2x25mlS8060-32P7: Standard/Sample Dilution Buffer, 1x25mlS8060-32P8: Secondary Antibody (Biotin)/Avidin (HRP) Dilution Buffer, 1x25mlS8060-32P9: TMB Substrate, 1x15mlS8060-32P10: Stop Solution (1N sulfuric acid), 1x15mlStorage and Stability:Store components at 4°C. Stable for 6 months. Aliquot and store *S8060-32P2 after reconstituted at -70°C. Standard can be thawed one time only without loss of immunoreactivity. For maximum recovery of product, centrifuge the original vial prior to removing the cap.