Human Sclerostin ELISA Kit

From Antibodies.com, this sandwich ELISA kit enables in vitro quantitative determination of human Sclerostin in serum, plasma, tissue homogenates, and other biological fluids. The format uses a pre-coated 96-well microplate (12 x 8 well strips) bearing an antibody specific for Sclerostin. Standards and samples are added to the wells, where Sclerostin is captured by the immobilized antibody. After washing, a Biotinylated Anti-Sclerostin Antibody is added, followed by HRP-Avidin conjugate. TMB substrate is then used to visualize the HRP enzymatic reaction, producing a blue-coloured product that turns yellow upon addition of acidic stop solution. Absorbance is read at 450 nm and the Sclerostin concentration is calculated against the standard curve.

The assay has a reported sensitivity of <0.122 ng/ml and a product range of 0.312–20 ng/ml, with a total assay time of approximately 4 hours 30 minutes. Detection is colorimetric on a microplate reader. Reported sample recovery averages are 91% for serum (n=5, range 80%–102%), 90% for EDTA plasma (n=5, range 81%–100%) and 84% for heparin plasma (n=5, range 80%–89%). Dilution linearity for 1:2, 1:4, 1:8 and 1:16 dilutions is reported as 87–91%, 87–107%, 74–101% and 92–97% for serum; 90–105%, 84–101%, 90–101% and 79–108% for EDTA plasma; and 84–95%, 92–105%, 82–105% and 89–91% for heparin plasma (all n=5).

Kit components include a pre-coated 96-well microplate (12 x 8 well strips), 2 vials of lyophilized standard, 120 µl of Detection Solution A, 120 µl of Detection Solution B, 20 ml of 30X Wash Buffer, 45 ml of Sample Dilution Buffer, 9 ml of TMB Substrate, 6 ml of Stop Solution, and 5 adhesive plate sealers. Standards, Detection Solutions and the Sample Dilution Buffer are stored at -20°C, while the Wash Buffer, TMB Substrate and Stop Solution are stored at +4°C. Kit components should not be used past their expiration date.