Description
Principle of the assay: The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the Interleukin 6 present in samples reacts with the anti-Interleukin 6 antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing, the Detection antibody, biotin conjugated anti-IL-6 is added and complexes are formed. Following a wash step, the horseradish peroxidase (HRP) conjugated Streptavidin is added and complexes are formed. After another washing step, the complexes are assayed by the addition of a chromo-genic substrate, 3,3',5,5'-tetramethylbenzidine (TMB). The quantity of bound enzyme varies directly with the concentration of IL-6 in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of IL-6 in the test sample. The quantity of IL-6 in the test sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution