Enzyme Immunoassay for the Quantitative Determination of Human Growth Hormone (HGH) in Human Serum
Principle of the assay: The Aviva HGH ELISA is based on the principle of a solid phase enzyme-linked immunosorbent assay (ELISA). The assay system utilizes a sheep anti-HGH antibody for solid phase (microtiter wells) immobilization and a mouse monoclonal anti-HGH antibody in the antibody-enzyme (horseradish peroxidase) conjugate solution. The test sample is allowed to react simultaneously with the antibodies, resulting in HGH 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 TMB is added and incubated for 20 minutes, resulting in the development of a blue color. The color development is stopped with the addition of 1N HCl, and the color is changed to yellow and measured spectrophotometrically at 450 nm. The concentration of HGH is directly proportional to the color intensity of the test sample.
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