Background |
The sCD27 ELISA kit is a sandwich-type of enzyme immunoassay in which a monoclonal CD27 antibody is coated to polystyrene microtiter wells. Soluble CD27 molecules present in a measured volume of sample or standard are bound by the antibodies on the microtiter plate, and non-bound material is removed by washing. Subsequently, a biotinylated second monoclonal CD27 antibody is added. This antibody binds to the sCD27 antibody complex present in the microtiter well. Excess biotinylated antibody is removed by washing, followed by addition of horseradish peroxidase (HRP) conjugated streptavidin, which binds onto the biotinylated side of the sCD27 antibody sandwich. After removal of non-bound streptavidine-HRP conjugate by washing, substrate solution is added to the wells. A coloured product is formed in proportion to the amount of sCD27 present in the sample or standard. After the reaction has been terminated by the addition of a stop solution, absorbance is measured in a microtiter plate reader. From the absorbance of samples and those of a standard curve, the concentration of sCD27 can be determined by interpolation with the standard curve. |
