Principle of Assay
Human Amyloid Precursor Protein ELISA Kit (A2148) employs the sandwich enzyme immunoassay technique for the quantitative measurement of human Amyloid Precursor Protein in serum, plasma, cerebrospinal fluid or other biological fluids. An antibody specific for Amyloid Precursor Protein has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the Amyloid Precursor Protein present in each sample is bound to the wells by the immobilized antibody. Following incubation, the wells are washed and then incubated with Biotinylated Anti-Amyloid Precursor Protein Antibody, which binds the captured Amyloid Precursor Protein present in each well. Following incubation, unbound biotinylated detection 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 proportional to the amount of Amyloid Precursor Protein captured in each well. The concentration of Amyloid Precursor Protein can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
