Growth Hormone (rat/mouse)

This assay is a Sandwich ELISA based, sequentially, on: 1) capture of rat or mouse Growth Hormone molecules from samples to the wells of a microtiter plate coated by a pre-titered amount of anti-Growth Hormone polyclonal antibodies, 2) wash away of unbound materials from samples, 3) binding of a second biotinylated anti-Growth Hormone polyclonal antibody to the captured molecules, 4) wash away of unbound materials from samples, 5) conjugation of horseradish peroxidase to the immobilized biotinylated antibodies, 6) wash away of free enzyme conjugates, and 7) quantification of immobilized antibody-enzyme conjugates by monitoring horseradish peroxidase activities in the presence of the substrate 3,3´,5,5´-tetramethylbenzidine. The enzyme activity is measured spectrophotometrically by the increased absorbency at 450 nm, corrected from the absorbency at 590 nm, after acidification of formed products.

Since the increase in absorbency is directly proportional to the amount of captured rat or mouse Growth Hormone in the unknown sample, the latter can be derived by interpolation from a reference curve generated in the same assay with reference standards of known concentrations of Rat Growth Hormone.