Principle of Assay
Leukotriene E4 ELISA Kit (A326987) employs the competitive enzyme immunoassay technique for the quantitative measurement of Leukotriene E4 in serum, plasma, and other biological fluids. The 96-well microtiter plate has been pre-coated with Leukotriene E4 antigen. During the incubation, Leukotriene E4 present in the samples or standards competes with the fixed amount of immobilized Leukotriene E4 for binding sites on the Biotinylated Anti-Leukotriene E4 Antibody. The more Leukotriene E4 present in a sample or standard, the less Biotinylated Anti-Leukotriene E4 Antibody that binds to the plate. Following incubation, unbound Biotinylated Anti-Leukotriene E4 Antibody is removed by washing, and an HRP-Streptavidin conjugate is added to the wells and the microtiter plate is incubated. Following incubation and washing, TMB substrate solution is then used to visualize the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow after addition of acidic stop solution. The density of yellow is inversely proportional to the amount of Leukotriene E4 present in each sample or standard. The concentration of Leukotriene E4 can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
