Description
Intended use The CA125 ELISA Kit is intended for the determination of the Cancer Antigen CA125 concentration in human serum. For research use only, not for use in diagnostic procedures.
Principle of the test The CA125 ELISA test is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay system utilizes a monoclonal antibody directed against a distinct antigenic determinant on the intact CA125 molecule is used for solid phase immobilization (on the microtiter wells). A rabbit anti-CA125 antibody conjugated to horseradish peroxidase (HRPO) is in the antibody-enzyme conjugate solution. The unknowns are allowed to react simultaneously with the two antibodies, resulting in the CA125 molecules being sandwiched between the solid phase and enzyme-linked antibodies. After 90 minutes of incubation at 37°C, the wells are washed with water to remove unbound labeled antibodies. A solution of TMB Reagent is added and incubated for 20 minutes, resulting in the development of a blue color. The color development is stopped with the addition of Stop Solution changing the color to yellow. The concentration of CA125 is directly proportional to the color intensity of the unknowns being tested. Absorbance is measured spectrophotometrically at 450 nm