Mouse Proteinase 3 ELISA Kit

A sandwich enzyme-linked immunosorbent assay (ELISA) kit from Antibodies.com is designed for the in vitro quantitative determination of mouse Proteinase 3 (PR3) in serum, plasma, tissue homogenates, or other biological fluids. The assay employs a 96-well microplate pre-coated with an antibody specific for PR3. Standards and samples are added to the wells, where PR3 is captured by the immobilised antibody; following incubation and washing, a biotinylated anti-PR3 antibody is added, followed by an HRP-avidin conjugate and TMB substrate. The enzymatic reaction produces a blue product that turns yellow upon addition of stop solution, and the concentration of PR3 is calculated from the absorbance read at 450 nm against a standard curve.

According to the manufacturer, the assay has a sensitivity of <0.058 ng/ml and a measuring range of 0.156–10 ng/ml, with a total assay time of 4h 30m. Spike-and-recovery performance was assessed in mouse samples (n=5): serum showed an average of 91% (range 80–102%), EDTA plasma an average of 90% (range 81–100%), and heparin plasma an average of 84% (range 80–89%). Dilution linearity was also evaluated at 1:2, 1:4, 1:8 and 1:16, yielding ranges of 87–107% for serum, 79–108% for EDTA plasma, and 82–105% for heparin plasma.

Kit components include a pre-coated 96-well microplate (12 x 8 well strips), lyophilised standard (2 vials), detection solutions A and B (120 µl each), 30X wash buffer (20 ml), sample dilution buffer (45 ml), TMB substrate (9 ml), stop solution (6 ml), and plate sealers (5 adhesive strips). Components are stored at either -20°C (microplate, standard, detection solutions, sample dilution buffer) or +4°C (wash buffer, TMB substrate, stop solution) as instructed.