Enzyme Immunoassay for the Quantitative Determination of Follicle-Stimulating Hormone (FSH) in Human Serum
Principle of the assay: The Aviva FSH EIA Test is based on the principle of a solid phase enzyme-linked immunosorbent assay.13,14 The assay system utilizes mouse monoclonal anti-alpha FSH for solid phase (microtiter wells) immobilization, and mouse monoclonal anti-beta FSH in the antibody-enzyme (horseradish peroxidase) conjugate solution. The test sample is allowed to react simultaneously with the antibodies, resulting in the FSH molecules being sandwiched between the solid phase and enzyme-linked antibodies. After a 45 minute incubation at room temperature, the wells are washed with water to remove unbound labeled antibodies. A solution of Tetramethylbenzidine (TMB) is added and incubated for 20 minutes at room temperature, resulting in the development of a blue color. The color development is stopped with the addition of 1N HCl, and the resulting yellow color is measured spectrophotometrically at 450 nm. The concentration of Follicle-Stimulating Hormone is directly proportional to the color intensity of the test sample.
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