• Specific antibodies targeting the receptor binding region
• Rigorously validated for clinical samples according to FDA/ICH/EMEA guidelines
• Low sample volume - 20 µl of serum/plasma per well 

In a first step, assay buffer is pipetted into the wells of the microtiter strips. Thereafter, standard/control/sample are pipetted into the wells, which are pre-coated with a recombinant monoclonal anti-human Sclerostin antibody. Any bioactive Sclerostin present in the standard/control/sample binds to the pre-coated antibody in the well. After incubation, a washing step is applied where all non-specific unbound material is removed. In a next step, the conjugate (anti-human Sclerostin-HRP) is pipetted into the wells and reacts with bioactive Sclerostin present in the sample, forming a sandwich. After another washing step, the substrate (tetramethylbenzidine; TMB) is pipetted into the wells. The enzyme-catalyzed color reaction of the substrate is directly proportional to the amount of bioactive sclerostin in the sample. This color change is detectable with a standard microtiter plate reader. A dose response curve of the absorbance (optical density, OD, at 450 nm) versus standard concentration is generated, using the values obtained from the standards. The concentration of bioactive Sclerostin in the sample is determined directly from the dose response curve.

Bioactive Sclerostin ELISA Typical Standard Curve

The figure below shows a typical standard curve for the bioactive human Sclerostin ELISA. The immunoassay is calibrated against recombinant human bioactive Sclerostin.

Bioactive Sclerostin ELISA Kit Components