Rat Sclerostin ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay designed for the in vitro quantitative determination of rat Sclerostin in serum, plasma, tissue homogenates, and other biological fluids. The Antibodies.com kit employs a pre-coated 96-well microtiter plate (12 x 8 well strips) on which an antibody specific for Sclerostin has been immobilized. Standards and samples are added to the wells and Sclerostin is captured by the immobilized antibody. After incubation and washing, a biotinylated anti-Sclerostin detection antibody is added, followed by an HRP-Avidin conjugate. TMB substrate is then used to visualize the enzymatic reaction, producing a blue product that turns yellow upon addition of acidic stop solution. Absorbance is read at 450 nm in a colorimetric microplate reader and the concentration of Sclerostin is calculated against the standard curve.

The assay has a sensitivity of <0.055 ng/ml and a product range of 0.156–10 ng/ml, with a total assay time of 4 hours 30 minutes. Kit components include a pre-coated 96-well microplate, lyophilized standard, detection solutions, wash buffer, sample dilution buffer, TMB substrate, stop solution, and plate sealers. Individual components are stored at either +4°C or -20°C as instructed in the protocol, and components should not be used past the indicated expiration date.

Manufacturer-reported spike-and-recovery values average 91% in serum (range 80–102%, n=5), 90% in EDTA plasma (range 81–100%, n=5), and 84% in heparin plasma (range 80–89%, n=5). Dilution linearity across 1:2, 1:4, 1:8, and 1:16 dilutions ranged from 87–91%, 87–107%, 74–101%, and 92–97% in serum; 90–105%, 84–101%, 90–101%, and 79–108% in EDTA plasma; and 84–95%, 92–105%, 82–105%, and 89–91% in heparin plasma, each with n=5.