Description
Aviva Systems Biology Inositol trisphosphate ELISA Kit (OKEH02606) is based on a competitive enzyme immunoassay technique. The microtiter well-plate in this kit has been pre-coated with an anti-Inositol trisphosphate antibody. Sample or standards are added to the wells along with a fixed quantity of biotinylated Inositol trisphosphate and incubated. The Inositol trisphosphate found in the sample or standards competes with the biotinylated Inositol trisphosphate for limited binding sites on the immobilized anti-Inositol trisphosphate antibody. Excess unbound biotinylated Inositol trisphosphate and sample or standard Inositol trisphosphate is washed from the plate. Avidin-HRP conjugate is added, incubated and washed. An enzymatic reaction is then produced through the addition of TMB substrate which is catalyzed by the immobilized HRP to generate a blue color product that changes yellow after adding acidic stop solution. The density of yellow coloration is measured by reading the absorbance at 450 nm which is quantitatively proportional to the amount of biotinylated Inositol trisphosphate captured in the well and inversely proportional to the amount of Inositol trisphosphate which was contained in the sample or standard