Description
This kit is a sandwich enzyme immunoassay for the quantitative determination of bioactive sclerostin in human serum and plasma samples (EDTA, citrate). In a first step, assay buffer is pipetted into the wells of the microtiter strips. Thereafter, STD/sample/CTRL are pipetted into the wells, which are pre-coated with the recombinant human monoclonal Sclerostin antibody. Any bioactive Sclerostin present in the STD/sample/CTRL 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 sclerostin-HRPO) is pipetted into the wells and reacts with bioactive Sclerostin present in the sample, forming a sandwich. After another washing step, the substrate (TMB Tetramethylbenzidine) is pipetted into the wells. The enzyme catalysed colour change of the substrate is directly proportional to the amount of bioactive sclerostin. This colour change is detectable with a standard microtiter plate ELISA 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