Introduction

Isoprostanes are prostaglandin-like compounds that are produced by free radical mediated peroxidation of lipoproteins. This kit is for the quantification of 15-isoprostane F_2t (also known as 8-epi-PGF_{2 a} or 8-iso-PGF_2a) in urine samples. Levels of 15-isoprostane F_2t in urine are useful for the non-invasive assessment of oxidant stress in vivo. 15-isoprostane F_2t 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 F_2t in urine serum samples without the need for prior purification or extraction.

Intended Use:

This kit is intended for the quantification of free 15-isoprostane F_2t the best characterized Isoprostane, in urine, without the need to extract the samples prior to testing.

NOTE: Plasma, tissue or cell lysates may be tested using the NWLSS NWK-ISO01 assay for isoprostanes. Detailed extraction procedures are included with the product.

Test Principle:

This assay utilizes a competitive enzyme-linked immunoassay (ELISA) strategy. Prior to assaying, all samples are treated with an Enhancing Reagent. After treatment with Enhancing Reagent, 15-isoprostane F2t in the samples or standards is allowed to compete with 15-isoprostane F_2t conjugated to horseradish peroxidase (HRP) for binding to a polyclonal antibody specific for 15-isoprostane F_2t coated on a microplate. Later TMB substrate addition results in a blue color development that is inversely proportional to the quantity of 15-isoprostane F_2t 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.