ree soluble RANKL ELISA highlights:

• High sensitivity – measurable concentrations in healthy subjects
• Only ELISA that measures free, uncomplexed soluble RANKL

RANKL ELISA Product Overview

The human RANKL ELISA kit is an overnight, 96-well sandwich ELISA for the quantitative determination of free soluble RANKL in serum and heparın plasma.

RANKL ELISA Assay Principle

The Biomedica RANKL ELISA is a sandwich enzyme immunoassay for the quantitative determination of free soluble RANKL in human serum and heparın plasma samples.

The figure below explains the principle of the RANKL human ELISA:

Target binding partner: recombinant human Osteoprotegerin (OPG)
Detection Antibody: polyclonal goat anti-human sRANKL
Target Antigen: free soluble human RANKL

In a first step, wells which are pre-coated with recombinant Osteoprotegerin (OPG), RANKL’s interaction partner, are prewashed to ensure optimal sensitivity. Thereafter, assay buffer is pipetted into the wells of the microtiter strips followed by the addition of standard/control/sample. Soluble RANKL present in the standard/control/sample binds to the pre-coated OPG in the well. After incubation, a the plate is washed to remove all non-specifically bound material. In a next step, biotinylated detection antibody (polyclonal goat anti-human sRANKL) is pipetted into the wells and reacts with the sRANKL present in the sample, forming a sandwich. Next, all unbound antibody is removed during another washing cycle. In the following step, the conjugate (streptavidin-polyHRP) is added and reacts with the detection antibody. After a final washing step, the substrate (TMB, tetramethylbenzidine) is pipetted into the wells. The enzyme-catalyzed color change of the substrate is directly proportional to the amount of soluble RANKL present 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 soluble RANKL in the sample is determined directly from the dose response curve.

RANKL ELISA Typical Standard Curve

The figure below shows a typical standard curve for the human RANKL ELISA kit. The immunoassay is calibrated against recombinant RANKL peptide:

RANKL ELISA Kit Components