Intended use
This immunoassay kit allows for the in vitro quantitative determination of General 12HETE concentrations in serum, Plasma, tissue homogenates and Cell culture supernates and Other biological fluids.
Test principle
The ELISA is based on the competitive binding enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with an antibody specific to General 12HETE, During the reaction, General 12HETE in the sample or standard competes with a fixed amount of biotin-labeled General 12HETE for sites on a pre-coated Monoclonal antibody specific to General 12HETE. Excess conjugate and unbound sample or standard are washed from the plate. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of General 12HETE in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 12-Hydroxyeicosatetraenoic acid and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

General Annotation

Function:

12-Hydroxyeicosatetraenoic acid (12-HETE) is an eicosanoid. It is a metabolite of arachidonic acid produced by the action of the enzyme arachidonate 12-lipoxygenase. 12-HETE is a highly selective ligand used to label mu opioid receptors.

Sequence:C20H32O3