Description
Aviva Systems Biology Hemoglobin ELISA Kit (Human) (OKCA00094) is based on a competitive enzyme immunoassay technique. The microtiter well-plate in this kit has been pre-coated with an anti-human Hemoglobin antibody. Sample or standards are added to the wells along with a fixed quantity of biotinylated Hemoglobin and incubated. The Hemoglobin found in the sample or standards competes with the biotinylated Hemoglobin for limited binding sites on the immobilized anti-human Hemoglobin antibody. Excess unbound biotinylated Hemoglobin and sample or standard Hemoglobin 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 Hemoglobin captured in the well and inversely proportional to the amount of Hemoglobin which was contained in the sample or standard