Isoprostane (8-iso-PGF2a) Serum, Tissue 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 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, plasma, 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. Principle: 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. Sensitivity: 0.1ng/ml Dynamic Range: 0.1-1.0ng/ml Performance Characteristics: Crossreactivity at 50% B/B0: 15-isoprostane F2t 100.0% 9a,11b-Prostaglandin F2a 4.1% 13,14-Dihydro-15-KETO-F2a 3.0% 9b,11a-Prostaglandin F2a <0.01% Prostaglandin F2a <0.01% 6-KETO-Prostaglandin F2a <0.01% Prostaglandin E2 <0.01% Prostaglandin D2 <0.01% Arachidonic Acid <0.01% Kit Components: 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 degrees C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.