The ImmunoDiangostics Insulin Immunoassay is a Solid Phase ELISA that is designed to accurately detect Insulin in human serum and plasma samples.
The ImmunoDiagnostics Insulin (ELISA) is a reliable and reproducible tool for the quantitative detection of Insulin antigen. The assay is intended for use on human serum or plasma.
Numerous application fields exist, but particularly:
- Diabetes: The inadequate production of insulin is a major cause of Type 2 Diabetes while the loss of insulin producing cells leading to insulin deficiency is the cause of Type 1 Diabetes.
Metabolic Syndrome: Insulin resistance has been proposed to be a basic underlying cause of the broad-based metabolic syndrome.
Significantly more affordable than competitor kits
Insulin Antibody Coated Microstrips: One microplate, 12 strips with 8 wells each, 96 dry wells in total. The wells are coated with a monoclonal anti- human Insulin antibody. The microplate is sealed in a foil bag. Any unused strips should be returned to the bag and resealed for future use.
Detection Antibody Solution: Concentrate (100x conc). One vial containing 0.12 mL of an HRP conjugated monoclonal anti- human Insulin. Detection antibody should be diluted with only the 1X Assay Buffer needed.
Assay Buffer: Concentrate (5x conc). One vial containing 20 mL of buffer for dilution of the detection antibody. If precipitates are observed in the concentrated buffer, warm at 37oC until the precipitates disappear. Dilute 20 mL of 5x Assay Buffer with 80 mL diH2O to make a 1x concentration prior to use.
Human Insulin Standards: 0 μIU/mL (5 mL). 3 μIU/mL, 6 μIU/mL, 12 μIU/mL, 25 μIU/mL, 50 μIU/mL, 120 μIU/mL (0.3 mL each). Ready to Use.
Wash Buffer: Concentrate (10x conc). One vial containing 50 mL of 10x wash buffer. If precipitates are observed in the 10x solution, warm at 37oC until the precipitates disappear. Prepare 1x Wash Buffer my mixing 50 mL of 10x Wash Buffer with 450 mL diH2O.
Substrate Solution: One bottle containing 12 mL of TMB (3,3’,5,5’-Tetramethylbenzidine) substrate. Ready-to-use. Do not expose to light.
Stop Solution: One vial containing 12 mL of ready-to-use stop solution.
The Insulin ELISA is a quantitative sandwich enzyme immunoassay. Standard, samples, and any desired controls are pipetted into the wells pre-coated with solid-phase capture antibody and co-incubated with an HRP conjugated monoclonal antibody targeting Insulin. After washing away any unbound substances a TMB solution is added to develop color in proportion to the amount of human Insulin initially bound. The color development is stopped by addition of an acidifying solution and the optical density is determined. By plotting a standard curve versus measured absorbance, the amount of antigen in the sample can be calculated. The concentration of the antigen is expressed in μIU/mL.
The kit contains calibrators and can be used in any laboratory equipped with a plate reader. The kit is optimized for the quantitative determination of human Insulin concentrations in serum plasma, and cell culture supernatant samples. The test can provide results within 90 minutes.
Example only! Do not use for calculations
4 Parameter Curve
Log-log Regression Curve
Considered to be the primary anabolic hormone, insulin is produced in the beta cells of the pancreas. Its main function is to regulate metabolism by promoting the absorption of glucose into the liver, muscles, and fat from the bloodstream. When the amount of glucose in the blood—known as blood sugar—is high, beta cells react by secreting insulin into the blood. By contrast, when blood sugar levels are low, insulin secretion is inhibited.
Animals with Type 1 diabetes have had their insulin producing beta cells destroyed in an autoimmune reaction. The result is an abnormally high blood sugar content and a generalized wasting away of the body. Those with Type 2 diabetes have lost beta functionality via accumulation of amyloid in the pancreatic islets compounded by insulin resistance in peripheral tissue. Often time multiple comorbidities can be observed, such as, hypertension, obesity, and cardiovascular disease in a poorly understood metabolic syndrome.
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