Description
This ELISA is designed for the quantitative determination of osteonectin. The test principle is based on a competition between antigen in the sample or standards and the antigen coated on the wells of microplate. Standards or samples are transferred with the primary antibody against osteonectin directly into the precoated microtiter plate. The antigen in the samples competes with the antigen immobilised on the microtiter plate for the free binding site of the specific antibodies against osteonectin. A peroxidase-conjugated antibody is used for detection and quantification, and tetramethylbenzidine (TMB) as a peroxidase substrate. The enzymatic reaction is terminated by acidic stop solution. A dose response curve of absorbance unit (optical density, OD at 450 nm) vs. concentration is generated using the values obtained from standard. The colour change is inversely proportional to the amount of analyte (sample or control). Osteonectin, present in the samples is determined directly from this curve