Human Osteopontin ELISA Kit

From Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (sELISA) is designed for the in vitro quantitative determination of human Osteopontin in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. A 96-well microtiter plate is pre-coated with an antibody specific for Osteopontin, and standards and samples are added so that the target is captured by the immobilised antibody. After washing, a Biotinylated Anti-Osteopontin Antibody binds the captured analyte, followed by an HRP-Avidin conjugate and TMB substrate. The resulting blue product turns yellow upon addition of acidic stop solution, and the concentration of Osteopontin is calculated from the absorbance at 450 nm read on a colorimetric microplate reader against the standard curve.

The kit has a product range of 0.625–40 ng/ml and a sensitivity of <0.234 ng/ml, with a total assay time of 4 hours 30 minutes. Reported average recoveries are 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 has been validated across serum, EDTA plasma and heparin plasma. The pre-coated 96-well microplate is supplied with lyophilised standard, Detection Solutions A and B, 30X wash buffer, sample dilution buffer, TMB substrate, stop solution and plate sealers, with kit components to be stored at +4 °C or −20 °C as instructed.