Rabbit Osteopontin ELISA Kit

Antibodies.com supplies a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) intended for the in vitro quantitative determination of rabbit Osteopontin in serum, plasma, tissue homogenates, or other biological fluids. The assay uses an antibody specific for Osteopontin that has been pre-coated onto a 96-well microtiter plate. Standards and test samples are added to the wells, allowing Osteopontin in each sample to be captured by the immobilised antibody. After washing, a Biotinylated Anti-Osteopontin Antibody binds the captured analyte, and an HRP-Avidin conjugate is added. TMB substrate solution is then used to visualise the HRP enzymatic reaction, producing a blue-coloured product that changes to yellow upon addition of acidic stop solution. The density of yellow is proportional to the amount of Osteopontin captured, and the concentration is calculated by reading the O.D. absorbance at 450 nm in a colorimetric microplate reader against the standard curve.

Performed as a quantitative colourimetric sandwich ELISA on a pre-coated microplate of 12 x 8 well strips, the kit has a sensitivity of <0.065 ng/ml and an assay range of 0.156–10 ng/ml, with a total assay time of 4h 30m. Spiked recovery was 80–102% (average 91%) in serum (n=5), 81–100% (average 90%) in EDTA plasma (n=5), and 80–89% (average 84%) in Heparin plasma (n=5). Dilution linearity was demonstrated across 1:2, 1:4, 1:8, and 1:16 dilutions for serum (87–91%, 87–107%, 74–101%, 92–97%), EDTA plasma (90–105%, 84–101%, 90–101%, 79–108%), and Heparin plasma (84–95%, 92–105%, 82–105%, 89–91%). Kit components should be stored at +4°C or -20°C as instructed and the kit must not be used past its expiration date.