The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to S100 Calcium Binding Protein (S100). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to S100 Calcium Binding Protein (S100). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain S100 Calcium Binding Protein (S100), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of S100 Calcium Binding Protein (S100) in the samples is then determined by comparing the O.D. of the samples to the standard curve.