Osteocalcin (1-43/49) Specific ELISA

This ELISA (enzyme-linked immunosorbent assay) kit is intended for the quantitative determination of both human osteocalcin (1-49) and osteocalcin (1-43) (also referred as N-terminal & mid-regional osteocalcin) levels in test samples. For research use only. Not for use in diagnostic procedures.

Test principle

This ELISA is designed, developed and produced for the quantitative measurement of human osteocalcin (1-49) and (1-43) in serum or plasma sample. The assay utilizes the two-site “sandwich” technique with two selected antibodies that bind to different epitopes of human osteocalcin.
Assay standards, controls and samples are added directly to wells of a microtiter plate that is coated with streptavidin. Subsequently, a mixture of biotinylated human osteocalcin N-terminal region specific polyclonal antibody and a peroxidase labeled human osteocalcin 20 – 43 region specific monoclonal antibody is added to each well. After the first incubation period, a “sandwich” of “biotinylated antibody - human osteocalcin – HRP-monoclonal antibody” is formed and this immunocomplex is also captured to the wall of microtiter plate via a streptavidin-biotin affinity binding. The unbound monoclonal antibodies and buffer matrix are removed in the subsequent washing step. A substrate solution in a timed reaction is then measured in a spectrophotometric microplate reader. The enzymatic activity of the immunocomplex bound to the wall of each microtiter well is directly proportional to the amount of human osteocalcin in a test sample. A standard curve is generated by plotting the absorbance versus the respective human osteocalcin concentration for each standard on point-to-point or 4 parameter curve fit. The concentration of human osteocalcin in test samples is determined directly from this standard curve.