Principle of Assay
Human 15-HETE ELISA Kit (A302944) is a Competitive ELISA for the quantitative measurement of human 15-HETE in serum, plasma, tissue homogenates, and other biological fluids. An antibody specific for 15-HETE has been pre-coated onto a 96-well microtiter plate. During the incubation, 15-HETE in the sample or standard competes with a fixed amount of Biotinylated-15-HETE. Following incubation, Biotinylated-15-HETE and free 15-HETE in the sample or standard are washed from the plate. HRP-Streptavidin (SABC) is then added to each well and incubated. Unbound SABC is washed away with wash buffer. TMB substrate solution is added and after incubation the enzyme-substrate reaction is terminated by the addition of sulfuric acid stop solution. The colour change is measured spectrophotometrically at a wavelength of 450nm and the concentration of 15-HETE in the samples is then determined by comparing the O.D. 450nm of the samples to the standard curve. O.D. 450nm is inversely proportional to the concentration of 15-HETE in the sample.
