Introduction
Isoprostanes are prostaglandin-like compounds that are produced by free radical mediated peroxidation of lipoproteins. Levels of 8-Isoprostane (15-isoprostane F2t), a representative isoprostane in biological fluids, have been shown to be useful for assessment of oxidant stress in vivo. 8-Isoprostane (15-isoprostane F2t) has also been shown to be a potent vasoconstrictor in rat kidneys and rabbit lungs and may play a causative role in atherogenesis. Elevated 8-isoprostane (15-isoprostane F2t) levels are associated with hepatorenal syndrome, rheumatoid arthritis, athero-sclerosis, and carcinogenesis.
Test Principle
This assay utilizes a competitive enzyme-linked immunoassay (ELISA) format. Briefly, 8-Isoprostane (15-isoprostane F2t) in the samples or standards is allowed to compete with 8-Isoprostane (15-isoprostane F2t) conjugated to horseradish peroxidase (HRP) for binding to a polyclonal antibody specific for 8-Isoprostane (15-isoprostane F2t) coated on a microplate. Subsequent TMB substrate addition results in a blue color development that is inversely proportional to the quantity of 8-Isoprostane (15-isoprostane F2t) in the original samples or standards. Addition of an acid stop solution causes a color change to yellow where absorbance is read at 450 nM.
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