The Gliadin IgG ELISA Assay Kit is an enzyme immunoassay for the quantitative determination of IgG antibodies to deamidated gliadin. The antibodies of the calibrators, positive control, and diluted samples react with fragments of deamidated gliadin immobilized on the solid phase of microtiter plates. After 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 the anti-human-IgG-antibodies conjugated to the 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. Evaluating the test by a semi-quantitative method is also possible. Sample Types: serum. Range: 1 - 300 U/ml. Sensitivity: 1 U/ml. Time: 2 hours ? The Eagle Biosciences Gliadin IgG ELISA Assay Kit is used for the quantitative determination of IgG antibodies against deamidated gliadin in human serum. The Gliadin IgG ELISA Assay Kit is for research use only and should not be used in diagnostic procedures.