Insulin ELISA

B-Bridge's Insulin ELISA kit is designed to quantitatively measure insulin present in human plasma and serum, and tissue culture medium. Due to the highly conserved nature of insulin it is expected that this kit will measure bovine and porcine insulin too.The human insulin protein is a 51 amino acid anabolic peptide-hormone that is secreted by the pancreatic beta-cells in the Islets of Langerhans. One of its primary functions is the stimulation of glucose uptake from the systemic circulation, as well as the suppression of hepatic gluconeogenesis, thereby serving a major role in glucose homeostasis and preventing the metabolic disorder diabetes mellitus. In addition to its role in regulating glucose metabolism, insulin stimulates lipogenesis, diminishes lipolysis, and increases amino acid transport into cells.Assay Principle:An Insulin standard 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 detected in a microtiter plate reader capable of measuring 450 nm wavelength. The concentration of the Insulin in the sample is calculated, after making suitable correction for dilution, using software available with most plate readers.

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