Assay Information
This SCC antigen EIA is a solidphase, noncompetitive immunoassay based upon the direct sandwich technique. Calibrators and samples are incubated together with biotinylated AntiSCC monoclonal antibody and horseradish peroxidase (HRP) labelled AntiSCC monoclonal antibody in Streptavidin coated microstrips. After washing, buffered Substrate/ Chromogen reagent (hydrogen peroxide and 3, 3’, 5, 5’ tetramethylbenzidine) is added to each well and the enzyme reaction is allowed to proceed. During the enzyme reaction a blue colour will develop if antigen is present. The intensity of the colour is proportional to the amount of SCC present in the samples. The colour intensity is determined in a microplate spectrophotometer at 620 nm (or optionally at 405 nm after addition of Stop Solution). Calibration curves are constructed for each assay by plotting absorbance value versus the concentration for each calibrator. The SCC concentrations of samples are then read from the calibration curve.