Indirect method, total duration of assay: 110 minutes.

The DiaSino Anti-TPO ELISA employs solid phase, indirect ELISA method for detection of antibodies to TPO in two-step incubation procedure. Polystyrene microwell strips are pre-coated with highly immunoreactive human TPO antigens. During the first incubation step, anti-TPO specific antibodies, if present, will be bound to the solid phase pre-coated TPO antigens. The wells are washed to remove unbound serum proteins, and rabbit anti-human IgG antibodies (anti-IgG) conjugated to the enzyme horseradish peroxidase (HRP-Conjugate) are added. During the second incubation step, these HRP-conjugated antibodies will be bound to any antigen-antibody(IgG) complexes previously formed and the unbound HRP-conjugate is then removed by washing.Chromogen solutions containing Tetramethylbenzidine (TMB) and urea peroxide are added to the wells and in presence of the antigen-antibody-anti-IgG (HRP) immunocomplex, the colorless Chromogens are hydrolyzed by the bound HRP conjugate to a bluecolored product. The blue color turns yellow after stopping the reaction with sulfuric acid. The amount of color intensity can be measured and it is proportional to the amount of antibody cap