Principle of Assay
Human APOA4 ELISA Kit (A119681) employs the sandwich enzyme immunoassay technique for the quantitative measurement of human APOA4 in serum, plasma, tissue homogenates, cell culture supernates and other biological fluids. An antibody specific for APOA4 has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the APOA4 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-APOA4 Antibody, which binds the captured APOA4 present in each well. Following incubation, unbound biotinylated detection antibody is removed by washing, and an HRP-Streptavidin 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 APOA4 captured in each well. The concentration of APOA4 can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
