Isoprostane (8-iso-PGF2a) Urine ELISA Kit, BioAssay(TM)

Isoprostanes are prostaglandin-like compounds that are produced by free radical mediated peroxidation of lipoproteins. This kit is for the quantification of 15-isoprostane F2t (also known as 8-epi-PGF2a or 8-iso-PGF2a) in urine samples. Levels of 15-isoprostane F2t in urine are useful for the non-invasive assessment of oxidant stress in vivo. 15-isoprostane F2t has also been shown to be a potent vasoconstrictor in rat kidneys and rabbit lungs, and plays a causative role in atherogenesis. Elevated isoprostane levels are associated with hepatorenal syndrome, rheumatoid arthritis, atherosclerosis, and carcinogenesis. This kit can be used for the quantification of free 15-isoprostane F2t in urine serum samples without the need for prior purification or extraction.\n\nPrinciple:\nThis kit is a competitive ELISA for determining levels of 15-isoprostane F2t (the best characterized isoprostane) in urine samples. Briefly, urine samples are mixed with an enhancing reagent that essentially eliminates interferences due to non-specific binding, and the 15-isoprostane F2t in the samples or standards competes with 15-isoprostane F2t conjugated to HRP (HRP) for binding to a polyclonal antibody specific for 15-isoprostane F2t coated on the microplate. The HRP activity results in color development when substrate is added, with the intensity of the color proportional to the amount of 15-isoprostane F2t bound and inversely proportional to the amount of unconjugated 15-isoprostane F2t in the samples or standards.\n\n\nKit Components: \nI9055-01: Microtiter plate precoated with 15-isoprostane F2t 1x96wells\nI9055-02: 15-isoprostane F2t Standard 2 x 60ul\nI9055-03: Enhancer Reagent 1x100ml\nI9055-04: Wash Buffer (5X) 1x40ml\nI9055-05: TMB Substrate 1x25ml\nI9055-06: 15-isoprostane F2t (HRP) 1x250ul\n\nStorage and Stability:\nStore at 4 degrees C. Do not freeze. Aliquots are stable for at least 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

I9055

Size

1Kit

Applications

ELISA

References

1. Morrow, J., Harris, T., & Roberts, L., Anal. Biochem. 14:1-10 (1990). 2. Morrow, J. D., Hill, K. , Burke, R. F., Nammour, T. M., Badr, K. F., and Roberts, L. J., Proc. Natl. Acad. Sci. U.S.A. 87:9383-9387 (1990). 3. Morrow, et. al., The Isoprostanes: Unique Bioactive Products of Lipid Peroxidation. Prog. Lipid Res. 36:1-21, 1997. 4. Morrow, J., Awad, J. A., Boss, H. J., Blair, I. A., and Roberts, L. J., Proc. Natl. Acad. Sci USA 89:10721-10725 (1992). 5. Wang, et al., Immunological characterization of urinary 8-epi-prostaglandin F2a excretion in man. J. Pharm. Exp. Ther. 275:94-100 (1995). 6. Morrow, J., Zackert, W., Yang, J., Kuhrts, E., Callewaert, D., Taber, D., Oates, J., Roberts, J., Quantitiation of the major urinary metabolite of the isoprostane 15-F2t-Isoprostane (8-iso-PGF2a) by a stable isotope dilution mass spectrometric assay, Analytical Biochem. 269: 326-331 (1999). 7. Roberts II, L.J., Morrow, J.D., Measurement of F2-isoprostanes as an index of oxidative stress in vivo, Free Radic. Biol. Med. 28:505-513 (2000).