Bovine Macrophage Inflammatory Protein 1 Alpha ELISA Kit

From Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (sELISA) is designed for the in vitro quantitative determination of bovine MIP-1 alpha/CCL3 in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids. The assay employs the sandwich enzyme immunoassay technique, in which an antibody specific for MIP-1 alpha/CCL3 is pre-coated onto a 96-well microtiter plate. Standards and test samples are added into the wells and the analyte present in each sample is bound by the immobilized antibody. After washing, a Biotinylated Anti-MIP-1 alpha/CCL3 Antibody binds the captured analyte, followed by addition of an HRP-Avidin conjugate. TMB substrate solution is then used to visualize the HRP enzymatic reaction, producing a blue-coloured product that changes to yellow after addition of acidic stop solution. The concentration of MIP-1 alpha/CCL3 is calculated by reading the O.D. absorbance at 450 nm in a microplate reader and referring to the standard curve.

The quantitative assay has a working range of 15.6–1000 pg/ml and a sensitivity of <7.4 pg/ml. Total assay time is 4 hours 30 minutes, and the platform is a pre-coated microplate of 12 x 8 well strips. Reported recovery values are: Serum 80–102% (average 91%, n=5), EDTA Plasma 81–100% (average 90%, n=5) and Heparin Plasma 80–89% (average 84%, n=5). Dilution linearity for serum (n=5) at 1:2, 1:4, 1:8 and 1:16 dilutions is 87–91%, 87–107%, 74–101% and 92–97% respectively; EDTA plasma gives 90–105%, 84–101%, 90–101% and 79–108%; and Heparin plasma gives 84–95%, 92–105%, 82–105% and 89–91%. Kit components should be stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date.