96 Wells
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Product Name Insulin
# of Samples 1 x 96 Assays.
Intended Use Direct immunoenzymatic colorimetric method for quantitative determination of Insulin in human serum or plasma.
Introduction Insulin is a polypeptide hormone that regulates carbohydrate metabolism. Apart from being the primary effector in carbohydrate homeostasis, it has effects on fat metabolism and it can change the liver's ability to release fat stores.
Insulin is involved in: control of cellular intake of glucose in muscle and adipose tissue, increase of DNA replication and protein synthesis, modification of the activity of numerous enzyme (allosteric effect), increased glycogen, fatty acid synthesis, amino acid uptake, decreased proteinolysis, lipolysis, gluconeogenesis.
Beta cells release insulin in a glucose-dependent way.
In most humans blood glucose levels varies from about 70 mg/dl to perhaps 110 mg/dl (3.9 to 6.1 mmol/l) except shortly after eating when the blood glucose level rises temporarily. This homeostatic effect is the result of many factors, of which hormone regulation is the most important.
There are several conditions in which insulin disturbance is pathologic: diabetes mellitus, insulinoma, metabolic syndrome and polycystic ovary syndrome. There are two types of diabetes mellitus: type 1 (autoimmune-mediated destruction of insulin producing beta cells in the pancreas resulting in absolute insulin deficiency), and type 2 (multifactor syndrome with combined influence of genetic susceptibility and influence of environmental factors, the best known being obesity, age, and physical inactivity, resulting in insulin resistance in cells requiring insulin for glucose absorption. This form of diabetes is strongly inherited). In both cases the insulin production must be increased by medication or delivering insulin by oral or by intravenous method.
The quantitative determination of insulin can help to determinate the dose to delivery.
Principles of the assay The Insulin ELISA test is based on simultaneous binding of human insulin by two monoclonal antibodies, one immobilized on microwell plates and the other conjugates with horseradish peroxidase (HRP).
After incubation, the bound/free separation is performed by a simple solid-phase washing, then the TMB-Substrate solution (TMB) is added. After an appropriate time has elapsed for maximum colour development, the enzyme reaction is stopped and the absorbancies are determinated.
The insulin concentration in the sample is calculated based on a series of standard.
The colour intensity is proportional to the insulin concentration in the sample.
Storage and Stability The closed reagents are stable up to the expiry date stated on the label when stored at 2-8°C in the dark.
Opened reagents are stable for 60 days when stored at 2-8°C.
Limitations of the Test Bacterial contamination or repeated freeze-thaw cycles of the specimen may affect the absorbance values.
References Eastham R.D: Biochemical Values in Clinical Medicine, 7th Ed. Bristol. England. Jonh Wright &Sons, Ltd; (1985).Gerbitz, VKD., Pankreatische B-Zellen Peptide: Kinetik und Konzentration von Proinsulin, Insulin und C- Peptid in Plasma and Urin. Probleme der Messmethoden. Klinische und Literaturübersicht. J. Clin. Chem. Biochem. 18, 313-326 (1980)Boehm TM, Lebovitz HE, Statistical analysis of Glucose and Insulin responses to intravenous tolbutamide: evaluation of hypoglycemic and hyperinsulinemic states: Diabetes Care 479-490 (1079)Wayne P A, National Committee for Clinical Laboratory Standards. Procedure for the collection of diagnostic blood specimens by venipuncture: approved standards. 4th Ed. NCCLS Document H3-A4, Wayne, PA(1988).Turkinton RW, Estkowski A, Link M, Secretion of Insulin dependence of connecting peptide; a predictor of insulin or dependence of obese diabetics. Archive of Internal Med. 142 (1982)Sacks BD, Carbohydrates in :Burtis, C.A. and Ashwood, AR (Eds) Tietz Textbook of Clinical Chemistry. 2nd Ed. Philadelphia W.B. Saunders Co. (1994)Kahn CR, Rosenthal AS, Immunologic reactions to insulin, insulin allergy, insulin resistance and autoimmune insulin syndrome. Diabetes Care 2, 283 295 (1979)

Additional Information

Name Insulin
Related Product Names Insulin
Molecular Weight 0.5
Storage The closed reagents are stable up to the expiry date stated on the label when stored at 2...8 °C in the dark.
Opened reagents are stable for 60 days when stored at 2…8°C.
Datasheets/Manuals Printable manual for GWB-9BC0DA
Intended Use Research Use Only