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I9055 Isoprostane (8-iso-PGF2a) Urine ELISA Kit, BioAssay™

Specifications
References
Brand
BioAssay™
Kit Type
ELISA
Tests
96
Detection Method
Colorimetric
Sample Matrix
Urine
EU Commodity Code
38220000
Shipping Temp
Blue Ice
Storage Temp
4°C

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.

Principle of the Assay
This 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 enhanced dilution buffer that essentially eliminates interference due to non-specific binding. The 15-isoprostane F2t in the samples or standards competes with 15-isoprostane F2t conjugated to 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.
Detection Range
0.05-100ng/ml
Kit Components
I9055-01: Microtiter Strip precoated with 15-isoprostane F2t, 1x96wells I9055-02: 15-isoprostane F2t Standard (1ug/ml), 2x60ul I9055-03: Enhanced Dilution Buffer 1x100ml I9055-04: Wash Buffer, 5X, 1x40ml I9055-05: TMB Substrate, 1x25ml I9055-06: 15-Isoprostane F2t (HRP), 1x250ul I9055-07: b-Glucuronidase, 2x100ul
Storage and Stability
Store other components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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).
USBio References
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