Total sRANKL ELISA

The Total sRANKL ELISA is intended for the quantitative determination of total sRANKL in human serum and plasma.

For research use only. Not for use in diagnostic procedures.

For Laboratory Professional Use Only.

Developed by Immundiagnostik AG.

Test Principle of the Total sRANKL ELISA

The Total sRANKL ELISA utilizes the two-site sandwich technique with two selected antibodies that bind to human sRANKL and OPG. Assay standards, controls, prediluted samples containing human sRANKL, and the OPG solutions are added to wells of a microplate coated with a high affine polyclonal anti-human OPG antibody.

After the first incubation period, sRANKL is bound to the OPG, and the antibody is immobilized on the wall of microtiter wells. Then a biotinylated monoclonal anti-human sRANKL antibody is added to each microtiter well, and a sandwich of capture antibody – human OPG – sRANKL – streptavidin (peroxidase-labeled) is formed.

For quantification, a streptavidin horseradish-peroxidase conjugate is added, which specifically binds to biotin. Tetramethylbenzidine (TMB) is used as a substrate for peroxidase. Finally, an acidic stop solution is added to terminate the reaction. The color changes from blue to yellow. The intensity of the yellow color is directly proportional to the concentration of total sRANKL.

A dose-response curve of the absorbance unit (optical density, OD at 450 nm) vs. concentration is generated using the values obtained from the standard. The presence of total sRANKL in the samples is determined directly from this curve.