Enzyme immunoassay for the direct, quantitative determination of melatonin in human saliva.
- The assay procedure follows the basic principle of competitive ELISA whereby there is competition between a biotinylated and a non-biotinylated antigen for a fixed number of antibody binding sites.
- The amount of biotinylated antigen bound to the antibody is inversely proportional to the analyte concentration of the sample. When the system is in equilibrium, the free biotinylated antigen is removed by a washing step and the antibody bound biotinylated antigen is determined by use of streptavidin-peroxidase as marker and TMB as substrate.
- Quantification of unknowns is achieved by comparing the enzymatic activity of unknowns with a response curve prepared by using known standards.