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Specifications
- Assay Type Sandwich ELISA
- Sample Types Serum, Plasma, Tissue Culture Media
- Sensitivity 48.8 pg/mL
- Species Human, Bovine, Porcine
- Assay Duration 2 Hours
- Samples/Plate 42 in Duplicate
- Readout Colorimetric, 450 nm
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Standard Curve
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Description
Assay Principle:
The Insulin ELISA Kit quantitatively measures Insulin levels in serum, plasma, and tissue culture media. The Insulin ELISA Kit is a sandwich ELISA with a run time of 2 hours. Please read the complete kit insert for more information before performing this assay.
Use our provided Human Insulin standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a transparent microtiter plate coated with our human Insulin monoclonal antibody and incubate covered shaking at room temperature for 1 hour. Add the peroxidase-conjugated human Insulin monoclonal antibody. Then incubate the mixture covered, shaking at room temperature for 30 minutes. The immunological reaction occurs between the peroxidase-conjugated Insulin antibody and the Insulin antigen in the sample or standard.
After the 30-minute incubation, wash away the excess peroxidase-conjugated Insulin antibody and add the TMB substrate. The TMB substrate reacts with the bound peroxidase-conjugated Insulin antibody generating a signal detected by a plate reader at 450nm. Use the intensity and the standard curve to calculate the Insulin concentration in the samples.
Background:
The human insulin protein is a 51 amino acid anabolic peptide-hormone secreted by the pancreatic β-cells in the Islets of Langerhans. Insulin consists of two chains (A and B) connected by disulfide bonds. One of its primary functions is the stimulation of glucose uptake from the systemic circulation and suppressing hepatic gluconeogenesis, thereby serving a significant role in glucose homeostasis and preventing the metabolic disorder diabetes mellitus.
The work of Banting, Best, Collip, and Macleod in the early 1920s resulted in identifying a substance in pancreas extracts that remarkably reduced blood glucose levels in diabetic animals. By 1923, physicians used these pancreatic extracts to treat diabetic patients successfully. Preproinsulin, the first translational product from the insulin gene, is a 110 amino acid polypeptide with a 24 amino acid signal peptide. Insulin exists primarily as a monomer at low concentrations(~10-6 M) and forms dimers at higher concentrations and neutral pH. At high concentrations and in the presence of zinc ions, insulin aggregates further to form hexameric complexes.
The primary function of insulin is to counter the concerted actions of several hyperglycemia-generating hormones and to maintain low blood glucose levels. In addition to regulating glucose metabolism, insulin stimulates lipogenesis, diminishes lipolysis, and increases amino acid transport into cells. Because there are numerous hyperglycemic hormones, untreated disorders associated with insulin generally lead to severe hyperglycemia and shortened life span.