Histamine ELISA

The Histamine ELISA is intended for the quantitative determination of histamine in EDTA plasma and urine.

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 8212.

Test Principle of the Histamine ELISA

The Histamine ELISA is designed for the quantitative determination of histamine in EDTA plasma or urine. The assay is based on the method of competitive enzyme-linked immunoassays.

The sample preparation includes the addition of a derivatization reagent for histamine derivatization. Afterward, the treated samples and a peroxidase-conjugated polyclonal histamine antibody are incubated in the wells of a microtiter plate coated with histamine derivative (tracer). During the incubation period, the target histamine in the sample competes with the tracer, immobilized on the wall of the microtiter wells, to bind the polyclonal 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 a photometer at 450 nm. The intensity of the yellow color is inversely proportional to the histamine concentration in the sample; this means high histamine concentration in the sample reduces the concentration of tracer-bound antibody 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 standards. The presence of histamine in the samples is determined directly from this curve.