Glutathione Peroxidase 1, Human, BioAssay(TM) ELISA Kit (GPX1, GPx-1, GSHPx-1, Cellular Glutathione Peroxidase)

Glutathione peroxidases (GPXs) are ubiquitously expressed proteins which catalyze the reduction of hydrogen peroxides and organic hydroperoxides by glutathione. There are several isoforms which differ in their primary structure and localization. The classical cytosolic /mitochondrial GPX1 (cGPX) is a selenium-dependent enzyme, first of the GPX family to be discovered. GPX2, also known as gastrointestinal GPX (GI-GPX), is an intracellular enzyme expressed only at the epithelium of the gastrointestinal tract. Extracellular plasma GPX (pGPX or GPX3) is mainly expressed by the kidney from where it is released into the blood circulation. Phospholipid hydroperoxide GPX4 (PH-GPX) expressed in most tissues, can reduce many hydroperoxides including hydroperoxides integrated in membranes, hydroperoxy lipids in low density lipoprotein or thymine. All mammalian GPX family members, except for the recently described Cys containing GPX3 and epididymis-specific secretory GPX (eGPX or GPX5) isoforms, possess selenocysteine at the active site. Because GPX1 appears to have a major role in the prevention of oxidative stress, it may also be an important antiatherogenic enzyme. In mice, GPX1 deficiency results in abnormal vascular and cardiac function and structure. From a clinical perspective, there is a report suggesting that low erythrocyte GPX1 activity identifies patients with coronary artery disease who are at the high risk for cardiovascular events and that measurement of GPX1 activity provides additional information on risk and might be useful in identifying patients who would benefit from preventive antioxidant treatment.\n\nIntended Use:\nHuman Glutathione Peroxidase-1 (human GPX1) ELISA kit is to be used for the in vitro quantitative determination of human GPX1 in human serum, human plasma, human RBC lysate, cell lysate and buffered solution. The assay will recognize both native and recombinant human GPX1. 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 GPX1 was calculated to be 1.56ng/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 crossreactivity: human GPX3, human GPX4, Rat GPX1 and mouse GPX1.\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 GPX1. Samples are pippetted into these wells. Nonbound GPX1 and other components of the sample should be removed by washing, then biotin-conjugated monoclonal antibody specific to GPX1 added. In order to quantitatively determine the amount of GPX1 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 GPX1.\n\nKit Components:\nG8127-11M1 - Microplate, 1x96 wells\nG8127-11M2 - Incubation Buffer, 1x30ml\nG8127-11M3 - Wash Buffer (20X), 2x25ml\nG8127-11M4 - Standard, 1x1 vial\nG8127-11M5 - Standard/Sample Dilution Buffer, 1x25ml\nG8127-11M6 - Secondary Antibody, 1x1 vial\nG8127-11M7 - Avidin-HRP (100X), 1x150ul\nG8127-11M8 - Secondary Ab/Avidin-HRP Dilution Buffer, 1x25ml\nG8127-11M9 - TMB Substrate, 1x15ml\nG8127-11M10 - Stop Solution, 1x15ml\n\nStorage and Stability:\nStore components at 4 degrees C. Any unused reconstituted standard should be discarded or frozen at -70 degrees C. Stable for 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

G8127-11M

Size

96Tests

References

1. Takebe, G., et al., J. Biol. Chem. 277(43): 41,254-41,258 (2002). 2. Avissar, N., et al., Am. J. Physiol. 267: E68-76 (1994). 3. Bao, Y., et al., FEBS Lett. 410: 210-212 (1997). 4. Chambers, I., et al., EMBO J. 5: 1221-1227 (1986). 5. Perry, A., et al., Biochem. J. 285: 863-870 (1992). 6. De Haan, J., et al., Redox Rep. 8(2): 69-79 (2003). 7. Blankenberg, S., et al., N. Eng. J. Med. 349: 1605-1613 (2003).