8-Isoprostane ELISA

This product is intended for the quantification of 15-isoprostane F2t (also known as 8-epi-PGF2a, 8-iso-PGF2a or more commonly 8-Isoprostane) and can be used with 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. NOTE: Alternatively, urine samples may be analyzed without solid phase extraction using the NWLSS™ NWK-ISO02 kit.

Introduction

Isoprostanes are prostaglandin-like compounds that are produced by free radical mediated peroxidation of lipoproteins. Levels of 15-isoprostane F_2t , a representative isoprostane in biological fluids, have been shown to be useful for 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, athero-sclerosis, and carcinogenesis.

Intended Use:

This kit is intended for the quantification of 15-isoprostane F_2t (also known as 8-epi-PGF_2a, 8-iso-PGF _2a or more commonly 8-Isoprostane) and can be used with 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 F_2t following hydrolysis of phospholipids.

NOTE: Alternatively, urine samples may be analyzed without solid phase extraction using the NWK-ISO2, Urinary Isoprostane kit.

Test Principle:

This assay utilizes a competitive enzyme-linked immunoassay (ELISA) format. Briefly, 15-isoprostane F_2t 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. Subsequent 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.

Assay Validation: