PRDX3, Human, BioAssay(TM) ELISA Kit (Thioredoxin-dependent Peroxide Reductase, Mitochondrial, Peroxiredoxin-3, Peroxiredoxin III, PRX III, Antioxidant Protein 1, AOP-1, Protein MER5 Homolog, HBC189, AOP1)

Organisms living under aerobic conditions have developed various anti-oxidative mechanisms to protect them from damage by reactive oxygen species (ROS). Peroxiredoxin (Prx) is a growing anti-oxidative protein family that has been identified six members in mammals. They share a common reactive Cys residue in the N-terminal region, and are capable of serving as a peroxidase and involve thioredoxin and/or glutathione as the electron donor. Prx1 to Prx4 have an additional Cys residue in the conserved C-terminal region, and are cross members as judged by the aa sequence similarity. Prx5 also contains an additional Cys in its C-terminal region that is less conserved. On the other hand, Prx6 has only one unique Cys. These Prx family members are distributed in subcellular localization, Prx1, 2, and 6 in cytosol, Prx3 in mitochondria, Prx4 in ER and secretion, Prx5 showing complicated distribution including peroxisome, mitochondria and cytosol, all of which are potential sites of ROS production. In addition to their role as a peroxidase, however, a body of evidence has accumulated to suggest that individual members also serve divergent functions, which are associated with various biological processes such as the detoxification of oxidants, cell proliferation, differentiation and gene expression. It would be expected that these functions might not necessarily depend on peroxidase activity and, therefore, it seems likely that the divergence is due to unique molecular characteristics intrinsic to each member. The specific localization of Prx3 in mitochondria together with the identification of its mitochondria-specific electron suppliers, namely thioredoxin 2 and thioredoxin reductase 2, suggest that these three proteins might provide a primary line of defense against H2O2 produced by the mitochondrial respiratory chain. Furthermore, Prx3 expression is induced by oxidants in the cardiovascular system and is thought to play a role in the antioxidant defense system and homeostasis within mitochondria.\n\nIntended Use:\nHuman Peroxiredoxin 3 (human Prx3) ELISA kit is to be used for the in vitro quantitative determination of human Prx3 in human mitochondrial fraction, human serum, cell lysate or buffered solution. The assay will recognize both native and recombinant human Prx3.\nThis 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 Prx3 was calculated to be 2ng/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.\n\nSpecificity:\nThe following substances were tested and found to have no cross-reactivity: human Prx1, Prx2, prx4, Prx5, Prx6, mouse Prx3 and rat Prx3.\n\nPrinciple:\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 Prx3. Samples are pippetted into these wells. Nonbound Prx3 and other components of the sample should be removed by washing, then biotin-conjugated monoclonal antibody specific to Prx3 added. In order to quantitatively determine the amount of Prx3 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. And 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 Prx3.\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

P3352-20M

Size

96Tests

Applications

ELISA

References

1. Sue Goo Rhee et al. Peroxiredoxins: a historical overview and speculative preview of novel mechanisms and emerging concepts in cell signaling. (2005) Free Radic Biol Med. 38(12):1543-52. 2. Fujii J, Ikeda Y. Advances in our understanding of peroxiredoxin, a multifunctional, mammalian redox protein. (2002) Redox Rep. 7(3):123-30. 3. Sue Goo Rhee et al. Peroxiredoxin, a novel family of peroxidases. (2001). IUBMB Life 52(1-2):35-41. 4. Tong-Shin Chang et al. Peroxiredoxin III, a mitochondrion-specific peroxidase, regulates apoptotic signaling by mitochondria. (2004) J Biol Chem. 279(40):41975-84. 5. Larisa Nonn et al. Increased expression of mitochondrial peroxiredoxin-3 (thioredoxin peroxidase-2) protects cancer cells against hypoxia and drug-induced hydrogen peroxide- dependent apoptosis. (2003) Mol Cancer Res. 1(9):682-9. 6. Zachary A. Wood et al. Structure, mechanism and regulation of peroxiredoxins. (2003) TIBS 28(1):32-40.