Isoprostane (8-iso-PGF2a) Serum, Tissue ELISA Kit, BioAssay™
|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 a representative isoprostane, 15-isoprostane F2t (also known as 8-epi-PGF2 or 8-iso-PGF2) Levels of 15-isoprostane F2t in biological fluids have been shown to be useful for 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 or tissue samples following solid phase extraction of the isoprostane-containing fraction. Instructions are also provided for the quantification of total 15-isoprostane F2t following hydrolysis of phospholipids.
This kit is a competitive ELISA for determining levels of 15-isoprostane F2t in biological samples. Briefly, 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 the samples or standards.The sensitivity of the assay is 0.1ng/ml, and the dynamic range is 0.1-1.0ng/ml.
I9050A: Microtiter Plate 1x96wells
I9050B: 15-isoprostane F2t Standard 2x60ul
I9050C: Wash Buffer (5X) 1x40ml
I9050D: Dilution Buffer (5X) 1x100ml
I9050E: Substrate 1x25ml
I9050F: 15-isoprostane F2t (HRP)1x100ul
Storage and Stability:
Store components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Dynamic Range: 0.1-1.0ng/ml
Cross reactivity at 50% B/B0:
15-isoprostane F2t 100.0%
9 ,11-Prostaglandin F2 4.1%
9 ,11-Prostaglandin F2 <0.01%
Prostaglandin F2 <0.01
6-KETO-Prostaglandin F1 <0.01%
Prostaglandin E2 <0.01%
Prostaglandin D2 <0.01%
Arachidonic Acid <0.01%
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 F2 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 Radical. Biol. Med. 28:505-513 (2000).