Human Carboxypeptidase A3, Mast Cell ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com, this Human CPA3 ELISA Kit is designed for the in vitro quantitative determination of human CPA3 in serum, plasma or other biological fluids. The assay employs a pre-coated 96-well microplate (12 x 8 well strips) onto which an antibody specific for CPA3 has been immobilised. Standards and test samples are added to the wells and any CPA3 present is captured by the immobilised antibody. After washing, a Biotinylated Anti-CPA3 Antibody is added, followed by an HRP-Avidin conjugate. A TMB substrate solution is then used to visualise the enzymatic reaction; the resulting blue product changes to yellow upon addition of an acidic stop solution. Absorbance is read at 450 nm in a microplate reader and the concentration of CPA3 is calculated by reference to a standard curve.

The assay has a quantitative range of 78.1-5000 pg/ml and a sensitivity of <34 pg/ml, with a total assay time of 4 hours 30 minutes. Validation data for serum and plasma samples report the following recovery ranges: serum (n=5) 80% - 102% (average 91%); EDTA plasma (n=5) 81% - 100% (average 90%); heparin plasma (n=5) 80% - 89% (average 84%). Reported linearity upon dilution at 1:2, 1:4, 1:8 and 1:16 is as follows: serum (n=5) 87-91%, 87-107%, 74-101%, 92-97%; EDTA plasma (n=5) 90-105%, 84-101%, 90-101%, 79-108%; heparin plasma (n=5) 84-95%, 92-105%, 82-105%, 89-91%. Kit components should be stored at +4°C or -20°C as instructed and not used past the expiration date.