Dopamine, ELISA

Dopamine is extracted by using a cis-diol-specific affinity gel, acylated and then derivatized enzymatically.

The competitive ELISA kit 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 concentrations 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 anti-rabbit 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.