Description
The Direct Salivary Melatonin EIA kit is based on the competition principal and microplate separation. Melatonin in standards (calibrators) and samples compete with a fixed amount of melatonin conjugated to horse radish peroxidase (Melatonin-HRP) for binding sites with a rabbit melatonin monoclonal antiserum bound to GARGG (goat anti-rabbit gamma globulin) coated wells of a microplate. After incubation, unbound components are washed away, enzyme substrate solution is added and a blue color formed. This reaction is stopped with an acid solution to produce a yellow color. The optical density is then read at 450 nm. The amount of Melatonin-HRP detected is inversely proportional to the amount of melatonin in a sample