Canine Macrophage Inflammatory Protein 1 Alpha ELISA Kit

This Antibodies.com assay is a sandwich ELISA (sELISA) intended for the in vitro quantitative determination of canine MIP-1 alpha/CCL3, also known as Macrophage Inflammatory Protein 1 Alpha, in serum, plasma or other biological fluids. The kit uses a pre-coated 96-well microplate arranged as 12 x 8 well strips. Standards and samples are added to the wells, where target analyte is captured by an immobilised specific antibody; bound analyte is then detected with a biotinylated anti-MIP-1 alpha/CCL3 antibody, followed by HRP-Avidin and TMB substrate. The enzymatic reaction produces a blue colour that turns yellow after stop solution is added, with absorbance read at 450 nm on a colorimetric microplate reader and concentration calculated from the standard curve.

Performance figures from the manufacturer state an assay range of 78.1-5000 pg/ml and a sensitivity of <28 pg/ml. The complete protocol runs in approximately 4h 30m. Spike-and-recovery across sample types (n=5 each) is reported as 80%-102% (average 91%) for serum, 81%-100% (average 90%) for EDTA plasma and 80%-89% (average 84%) for heparin plasma. Dilution linearity at 1:2, 1:4, 1:8 and 1:16 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. Kit components are stored at +4°C or -20°C as instructed, and the kit must not be used past its expiration date.