Human 8-iso prostaglandin F2α (8-iso-PGF2a) ELISA Kit

The isoprostanes are a family of eicosanoids of non-enzymatic origin produced by the random oxidation of tissue phospholipids by oxygen radicals. Isoprostanes appear as artifacts in tissue and plasma samples which have undergone oxidative degradation during prolonged or improper storage. They also appear in the plasma and urine under normal conditions and are elevated by oxidative stress. At least one of the isoprostanes, 8-isoprostane (8-epi PGF2α), has been shown to have biological activity. It is a potent pulmonary and renal vasoconstrictor1 and has been implicated as a causative mediator of hepatorenal syndrome and pulmonary oxygen toxicity.2 8-Isoprostane has been proposed as a marker of antioxidant deficiency and oxidative stress and elevated levels have been found in heavy smokers.3 8-Isoprostane levels are also a relative indicator of sample integrity for lipid-containing samples such as serum, plasma, and whole cell preparations.

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Human 8-iso prostaglandin F2α (8-iso-PGF2a) ELISA Kit has high sensitivity and excellent specificity for detection of Human 8-iso-PGF2a. No significant cross-reactivity or interference between Human 8-iso-PGF2a and analogues was observed.

This Human 8-iso prostaglandin F2α (8-iso-PGF2a) ELISA Kit employs a two-site sandwich ELISA to quantitate 8-iso-PGF2a in samples. An antibody specific for 8-iso-PGF2a has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any8-iso-PGF2a present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for 8-iso-PGF2a is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of 8-iso-PGF2a bound in the initial step. The color development is stopped and the intensity of the color is measured.

• Do not mix components from different kit lots or use reagents beyond the kit expiration date.• Allow all reagents to warm to room temperature for at least 30 minutes before opening.• Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.• Unused wells must be kept desiccated at 4 °C in the sealed bag provided.• Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.• It is recommended that all samples and standards be assayed in duplicate or triplicate.