SOD3, Human, BioAssay(TM) ELISA Kit (Extracellular Superoxide Dismutase Cu-Zn] EC-SOD)

Superoxide dismutase (SOD) is an antioxidative 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. Three unique and highly compartmentalized mammalian superoxide dismutases have been biochemically and molecularly characterized to date. SOD1, or CuZn-SOD (EC 1.15.1.1), was the first enzyme to be characterized and is a copper and zinc- containing homodimer that is found almost exclusively in intracellular cytoplasmic spaces. SOD2, or Mn-SOD (EC 1.15.1.1), exists as a tetramer and is initially synthesized containing a leader peptide, which targets this manganese-containing enzyme exclusively to the mitochondrial spaces. SOD3, or EC-SOD (EC 1.15.1.1), is the most recently characterized SOD, exists as a copper and zinc-containing tetramer. SOD3, which is highly expressed in selected tissues including blood vessels, heart, lungs, kidney and placenta, is found in the extracellular matrix of tissues and is ideally situated to prevent cell and tissue damage initiated by extracellularly produced ROS. SOD3 contains a unique heparin-binding domain at its carboxy-terminus that establishes localization to the extracellular matrix where the enzyme scavenges superoxide anion. SOD3 plays an important role in maintaining vascular tone, attenuating age-related cognitive decline, lung function, and the metabolism of NO, and in the pathology of such diseases as atherosclerosis, diabetes, and arthritis.\n\nIntended Use:\nHuman Superoxide Dismutase-3 (human SOD3) ELISA kit is to be used for the in vitro quantitative determination of human SOD3 in cell lysate and buffered solution. The assay will recognize both native and recombinant human SOD3. This kit has been configured for research use only and is not to be used in diagnostic procedures.\n\nSensitivity:\nThe minimal detectable dose of human SOD3 was calculated to be 2ng/ml, by subtracting\ntwo 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.\n\nSpecificity:\nThe following substances were tested and found to have no cross-reactivity: human SOD1, SOD2, SOD4.\n\nTest Principle:\nThe 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 SOD3. Samples are pippetted into these wells. Nonbound SOD3 and other components of the sample should be removed by washing, then biotin-conjugated monoclonal antibody specific to SOD3 added. In order to quantitatively determine the amount of SOD3 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 SOD3.\n\nKit Components:\nMicroplate: 1x96 wells\nIncubation buffer: 1x30ml\nWashing buffer, (10x): 1x100ml\nStandard protein: 1 glass vial lyophilized\nStandard/sample dilution buffer: 1x25ml\nSecond antibody: 1 glass vial lyophilized\nAV-HRP, (100x): 1x150ul\nSecondary antibody/AV-HRP dilution buffer: 1x25ml\nSubstrate (TMB): 1x20ml\nStop solution: 1x20ml\n\nStorage and Stability:\nStore components at 4 degrees C. Stable for at least 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

S8060-03M

Size

96Tests

References

1. Steen V. Petersen et al. The dual nature of human extracellular superoxide dismutase: One sequence and two structures (2003) PNAS 100(24):13875-13880. 2. Tim D. Oury et al. Human extracellular superoxide dismutase is a tetramer composed of two disulphide-linked dimmers: a simplified, high-yield purification of extracellular superoxide dismutase (1996) Biochem .J .317:51-57. 3. Tohru Fukai et al. Extracellular superoxide dismutase and cardiovascular disease (2002) Cardiovascular Research 55:239-249 4. Yoshiaki Furukawa et al. Oxygen-induced maturation of SOD1: a key role for disulfide formation by the copper chaperone CCS (2004) The EMBO Journal 23:2872-2881. 5. Richard W. Strange et al. The Structure of Holo and Metal-dificient Wild-type Human Cu, Zn Superoxide Dismutase and its Relevance to Familial Amyotrophic Lateral Sclerosis (2003) J. Mol. Biol. 328:877-891. 6. Stefan I. Liochev et al. Cross-compartment protection by SOD1 (2005) Free Radical Biology & Medicine 38:146-147. 7. Richard A. Weisiger et al. Superoxide Dismutase (1973) The Journal of Biol. Chem. 48(10):3582-3592. 8. Igor N. Zelko et al. Superoxide dismutase multigene family: a comparison of the CuZn- SOD (SOD1), Mn-SOD(SOD2), and EC-SOD(SOD3) gene structures, evolution, and expression (2002) Free Radical Biology & Medicine 33(3):337-349.