The DetectX Insulin EIA kit is designed to quantitatively measure insulin present in a variety samples and tissue culture media. An Insulin standard, calibrated to the WHO International Standard 1975 (66/304), is provided to generate a standard curve for the assay and all samples should be read off the standard curve. Standards or diluted samples are pipetted into a clear microtiter plate coated with a monoclonal antibody to capture insulin present in the sample. After a 60 minute incubation, the plate is washed. A peroxidase conjugated insulin antibody is added and the plate is again incubated for 30 minutes and washed. Substrate is then added to the plate, which reacts with the bound insulin conjugated antibody. After a third incubation, the reaction is stopped and the intensity of the generated color is read at 450 nm.