This CLIA kit operates on the Sandwich-CLIA principle. It comes with a micro CLIA plate pre-coated with an antibody specific to Human PAP. Samples or Standards are added to the plate wells, combined with the specific antibody. Then, a biotinylated detection antibody specific for Human PAP and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively, incubated, and free components are washed away. The substrate solution is added, and only wells containing Human PAP, biotinylated detection antibody, and Avidin-HRP conjugate exhibit luminescence. The Relative light unit (RLU) value is measured at 425nm, positively correlating with Human PAP concentration. You can calculate the concentration of Human PAP in samples by comparing the RLU value to the standard curve.