Description
The Eagle Biosciences Anti-hu tTG IgG ELISA Assay Kit is an enzyme immunoassay for the quantitative or semi-quantitative determination of IgG autoantibodies to tissue transglutaminase in human serum or plasma. Autoantibodies of the diluted patient samples, positive control, and calibrators react with human tissue transglutaminase immobilized on the solid phase of a microtiter plate. Anti-huTransG guarantees the specific binding of anti-tTG IgG autoantibodies of the specimen under investigation by employing highly purified, activated recombinant human tTG for coating. Following an incubation period of 60 min at room temperature (18…25°C), unbound serum components are removed by a wash step.The bound autoantibodies react specifically with anti-human-IgG-antibodies conjugated to horseradish peroxidase (HRP) within the incubation period of 30 min at room temperature. Excessive conjugate is separated from the solid-phase immune complexes by the following wash step.HRP converts the colorless substrate solution of 3,3’,5,5’-tetramethyl¬benzidine (TMB) added into a blue product. This enzyme reaction is stopped by dispensing an acidic solution (H2SO4) into the wells after 15 min at room temperature turning the solution from blue to yellow.The optical density (OD) of the solution at 450 nm is directly proportional to the amount of specific antibodies bound. The standard curve is established by plotting the concentrations of the antibodies of the calibrators (x-axis) and their corresponding OD values (y-axis) measured. The concentration of antibodies of the specimen is directly read off the standard curve