The Ferritin Quantitative Test is based on the principle of a solid phase enzyme-linked immuno sorbent assay.12,13,14The assay system utilizes rabbit anti-ferritin for the solid phase (microtiter wells) immobilization and mousemonoclonal anti-ferritin in the antibody-enzyme (horseradish peroxidase) conjugate solution. The test sample is allowed to react simultaneously with the antibodies, resulting in the ferritin 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 3,3’,5,5’-Tetramethylbenzidine (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 resulting yellow color is measured spectro photo metrically at 450 nm. The concentration of ferritin is directly proportional to the color intensity of the test sample.
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