Description
Principle of the Assay: This HBsAg ELISA kit uses polystyrene microwell strips pre-coated with monoclonal antibodies specific to HBsAg. Patient's serum or plasma sample is added to the microwell together with a second antibody conjugated with horseradish peroxidase (HRP) and directed against a different epitope of HBsAg. During incubation, the specific immunocomplex formed in case of presence of HBsAg in the sample, is captured on the solid phase. After washing to remove sample serum proteins and unbound HRPconjugate, Chromogen solutions containing tetramethylbenzidine (TMB) and urea peroxide are added to the wells. In presence of the antibody-antigenantibody(HRP) "sandwich" immunocomplex, the colorless Chromogens are hydrolyzed by the bound HRP-conjugate to a blue colored product. The blue color turns yellow after stopping the reaction with sulfuric acid. The amount of color can be measured and is proportional to the amount of antigen in the sample. Wells containing samples negative for HBsAg remain colorless.: