PrincipleThe micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human BMP-4. Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human BMP-4 and Avidin-Horseradish Peroxidase (HRP) conjugate is added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human BMP-4, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit(RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human BMP-4. You can calculate the concentration of Human BMP-4 in the samples by comparing the RLU value of the samples to the standard curve.