Osteopontin (OPN) ELISA

Osteopontin (OPN) ELISAThe Osteopontin ELISA is an enzyme immunoassay for the quantitative determination of Osteopontin in human serum and plasma. The microtiter plate is coated with the antibody specifically binding the Osteopontin. The human serum or plasma is incubated in the plate with the capture antibody. The specimen is washed out and the specifically bound protein is incubated with biotin labelled detection antibody. Following another washing step, Streptavidin HRP

The Osteopontin ELISA is an enzyme immunoassay for the quantitative determination of Osteopontin in human serum and plasma.

The microtiter plate is coated with the antibody specifically binding the Osteopontin. The human serum or plasma is incubated in the plate with the capture antibody. The specimen is washed out and the specifically bound protein is incubated with biotin-labelled detection antibody. Following another washing step, Streptavidin-HRP conjugate is added into the well. Unbound reagent is then washed out. Horseradish peroxidase (HRP) bound in the complex reacts with the chromogenic substrate (TMB) creating the blue colour. The reaction is stopped by addition of STOP solution (H2SO4). The absorbance values are measured at 450 nm (optionally 450/630 nm) and are proportional to the concentration of Osteopontin in the specimen. The concentration of Osteopontin in unknown samples is determined from the calibration curve which is created by plotting the absorbance values against the standard concentration values.