Human Chymase 1, Mast Cell ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com, designed for the in vitro quantitative determination of human Mast Cell Chymase in serum, plasma, tissue homogenates, cell lysates or other biological fluids. The assay employs the sandwich enzyme immunoassay technique: an antibody specific for Mast Cell Chymase is pre-coated onto the microplate, standards and test samples are added and the target is bound to the wells by the immobilised antibody. After washing, a Biotinylated Anti-Mast Cell Chymase Antibody is applied, followed by HRP-Avidin conjugate. Following further incubation and washing, TMB substrate is added; catalysis produces a blue-coloured product that changes to yellow upon addition of acidic stop solution. The density of yellow is proportional to the amount of Mast Cell Chymase captured in each well and is read as O.D. absorbance at 450 nm in a microplate reader, with the concentration calculated by reference to a standard curve. Kit components are to be stored at +4°C or -20°C as instructed and the kit should not be used past its expiration date.

The assay is quantitative and colourimetric, with a sensitivity of <6.5 pg/ml and a working range of 15.6–1000 pg/ml. Total assay time is 4h 30m and reactivity is human. Spiked recovery across five replicates averaged 91% in serum (range 80–102%), 90% in EDTA plasma (81–100%) and 84% in heparin plasma (80–89%). Linearity was demonstrated at dilutions of 1:2, 1:4, 1:8 and 1:16: serum recoveries of 87–91%, 87–107%, 74–101% and 92–97%; EDTA plasma recoveries of 90–105%, 84–101%, 90–101% and 79–108%; heparin plasma recoveries of 84–95%, 92–105%, 82–105% and 89–91%.