Technical Data
I9055
Isoprostane (8-iso-PGF2a) Urine ELISA Kit, BioAssay™
1Kit
Kits and Assays Storage: 4°C Do Not FreezeShipping: Blue Ice
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.

Principle:
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 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.


Kit Components:
I9055-01: Microtiter plate precoated with 15-isoprostane F2t 1x96wells
I9055-02: 15-isoprostane F2t Standard 2 x 60ul
I9055-03: Enhancer Reagent 1x100ml
I9055-04: Wash Buffer (5X) 1x40ml
I9055-05: TMB Substrate 1x25ml
I9055-06: 15-isoprostane F2t (HRP) 1x250ul

Storage and Stability:
Store at 4°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.
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
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).

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.