Mouse Alkaline Phosphatase, Liver/Bone/Kidney ELISA Kit

From Antibodies.com, this sandwich ELISA is configured for the in vitro quantitative determination of mouse Alkaline Phosphatase (Liver/Bone/Kidney) in serum, plasma, tissue homogenates and other biological fluids. The assay uses a pre-coated microplate supplied as 12 x 8 well strips and follows a sandwich enzyme immunoassay principle: analyte in standards and samples is captured by an immobilized antibody specific to the target, followed by detection with a biotinylated anti-Alkaline Phosphatase (Liver/Bone/Kidney) antibody and an HRP-Avidin conjugate. After addition of TMB substrate, the reaction yields a blue product that turns yellow upon addition of acidic stop solution; absorbance is read at 450 nm in a microplate reader and the target concentration is calculated against the standard curve.

Kit performance specifications include a measuring range of 0.156–10 ng/ml and a sensitivity of 0.057 ng/ml, with a total assay time of approximately 4 hours 30 minutes. Spike-and-recovery testing (n=5) gave the following results: serum 80–102% range with a 91% average, EDTA plasma 81–100% range with a 90% average, and heparin plasma 80–89% range with an 84% average. Dilution linearity was demonstrated at 1:2, 1:4, 1:8 and 1:16, with recoveries of 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, respectively. Provided reagents include the pre-coated microplate, lyophilized standard, detection solution A, detection solution B, wash buffer, sample dilution buffer, TMB substrate, stop solution, and plate sealers, with storage at +4°C or -20°C as indicated in the protocol.