Special remarks:

Intended use and principle of the test
Enzyme Immunoassay for the quantitative determination of 5-Hydroxy-3-Indole Acetic Acid (5-HIAA) in urine.

First, 5-HIAA is quantitatively derivatized by methylation. The subsequent competitive ELISA uses the microtiter plate format. The antigen is bound to the solid phase of the microtiter plate. The derivatized standards, controls and samples and the solid phase bound analyte compete for a fixed number of
antiserum binding sites.

After the system is in equilibrium, free antigen and free antigen-antiserum complexes are removed by washing. The antibody bound to the solid phase is detected by an antirabbit IgG-peroxidase conjugate using TMB as a substrate. The reaction is monitored at 450 nm.

Quantification of unknown samples is achieved by comparing their absorbance with a reference curve prepared with known standard concentrations.