The IDK® Serotonin ELISA is intended for the quantitative determination of serotonin (5-hydroxytryptamine, 5-HT) in human serum and dried blood spots.

For research use only. Not for use in diagnostic procedures.

For Laboratory Professional Use Only.

This is the RUO version of Immundiagnostik part number K 6880.

Test Principle of the IDK® Serotonin ELISA

The IDK® Serotonin ELISA is designed for the quantitative determination of serotonin in human serum and dried blood spots. The assay is based on the method of competitive enzyme-linked immunoassays. The sample preparation includes the addition of a derivatization reagent for serotonin derivatization. Afterward, the treated samples and the polyclonal serotonin antiserum are incubated in the wells of a microtiter plate coated with serotonin derivative (tracer).

During the incubation period, the target serotonin in the sample competes with the tracer immobilized on the wall of the microtiter wells to bind the polyclonal antibodies. During the second incubation step, a peroxidase-conjugated antibody is added to detect the anti-serotonin antibodies. After washing away the unbound components, tetramethylbenzidine (TMB) is added as a peroxidase substrate.

Finally, the enzymatic reaction is terminated by an acidic stop solution. The color changes from blue to yellow, and the absorbance is measured in the photometer at 450 nm. The intensity of the yellow color is inversely proportional to the serotonin concentration in the sample; this means a high serotonin concentration in the sample reduces the concentration of tracer-bound antibodies and lowers the photometric signal.

A dose-response curve of the absorbance unit (optical density, OD at 450 nm) vs. concentration is generated using the values obtained from the standard. Serotonin, present in the samples, is determined directly from this curve.