Principle of Assay
Human Tripeptidyl Peptidase II/TPPII ELISA Kit (A270841) employs the competitive enzyme immunoassay technique for the quantitative measurement of human Tripeptidyl Peptidase II/TPPII in serum, plasma, tissue homogenates or other biological fluids. The 96-well microtiter plate has been pre-coated with Tripeptidyl Peptidase II/TPPII antigen. During the incubation, Tripeptidyl Peptidase II/TPPII present in the samples or standards competes with the fixed amount of immobilized Tripeptidyl Peptidase II/TPPII for binding sites on the Biotinylated Anti-Tripeptidyl Peptidase II/TPPII Antibody. The more Tripeptidyl Peptidase II/TPPII present in a sample or standard, the less Biotinylated Anti-Tripeptidyl Peptidase II/TPPII Antibody that binds to the plate. Following incubation, unbound Biotinylated Anti-Tripeptidyl Peptidase II/TPPII Antibody is removed by washing, and an HRP-Avidin conjugate is added to the wells and the microtiter plate is incubated. Following incubation and washing, TMB substrate solution is then used to visualize the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow after addition of acidic stop solution. The density of yellow is inversely proportional to the amount of Tripeptidyl Peptidase II/TPPII present in each sample or standard. The concentration of Tripeptidyl Peptidase II/TPPII can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
