Description
The Eagle Biosciences Anti- ß2 GP-I ELISA Assay Kit is an enzyme immunoassay for the quantitative determination of IgG or IgM antibodies to ß2 glycoprotein-I in human serum or plasma. The antibodies of the standards, the positive control and diluted patient samples react with the human ß2 GP-I, immobilized on the solid phase of microtiter plates. The use of highly purified ß2 GP-I guarantees the specific binding of antibodies to ß2 glycoprotein-I of the specimen under investigation. Following an incubation period of 60 min at room temperature (RT), unbound serum components are removed by a wash step.The bound antibodies react specifically with anti-human-IgG or IgM conjugated to horseradish peroxidase (HRP) within the next incubation period of 30 min at RT. Excessive conjugate is separated from the solid-phase immune complexes by an additional wash step. HRP converts the colorless substrate solution of 3,3`,5,5`-tetramethyl-benzidine (TMB) added into a blue product. The enzyme reaction is stopped by dispensing an added acidic solution (H2SO4) into the wells, after 15 min at RT turning the solution from blue to yellow.The optical density (OD) of the solution at 450 nm is directly proportional to the amount of specifically bound antibodies.The standard curve is established by plotting the antibody concentrations of the standards (x-axis) and their corresponding OD-values (y-axis) measured. The concentration of antibodies of the specimen is directly read off the standard curve