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Canine Insulin EIA 狗胰島素檢測(cè)試劑盒背景介紹：

Mercodia Canine Insulin ELISA provides a method for the quantitative determination of insulin  in canine serum and plasma.

Canine Insulin EIA 狗胰島素檢測(cè)試劑盒SUMMARY AND EXPLANATION OF THE TEST

Insulin is the principal hormone responsible for the control of glucose metabolism. It is synthesised in the ?-cells of the islets of Langerhans as the precursor, proinsulin, which is processed toform C-peptide and insulin. Both are secreted in equimolar amounts into the portal circulation.The mature insulin molecule comprises two polypeptide chains, the A chain and the B chain. Thetwo chains are linked together by two inter-chain disulphide bridges. There is also an intra-chaindisulphide bridge in the A chain.Secretion of insulin is mainly controlled by plasma glucose concentration, and the hormonehas a number of important metabolic actions. Its principal function is to control the uptakeand utilisation of glucose in peripheral tissues via the glucose transporter. This and other hypoglycaemic activities, such as the inhibition of hepatic gluconeogenesis and glycogenolysis arecounteracted by the hyperglycaemic hormones including glucagon, epinephrine (adrenaline),growth hormone and cortisol.Diabetes mellitus is one of the most common endocrine disorders in dogs, with clinical features resembling the human type 1 diabetes, such as polydipsia, polyuria and weight loss (1-2).Diabetes is diagnosed in dogs with persistant hyperglycaemia (>9 mmol/L) and glucosuria. Diabetes typically occurs in dogs between 5 to 12 years of age. Furthermore, certain breeds of dogappear to be predisposed to diabetes and female dogs are more likely to develop diabetes (3).

Canine Insulin EIA 狗胰島素檢測(cè)試劑盒PRINCIPLE OF THE PROCEDURE

Mercodia Canine Insulin ELISA is a solid phase two-site enzyme immunoassay. It is based on thedirect sandwich technique in which two monoclonal antibodies are directed against separateantigenic determinants on the insulin molecule. During incubation, insulin in the sample reactswith peroxidase-conjugated anti-insulin antibodies and anti-insulin antibodies bound to themicroplate. After a simple washing step that removes unbound enzyme labelled antibody,the bound conjugate is detected by reaction with 3,3′-5,5′-tetramethylbenzidine (TMB). Thereaction is stopped by the addition of acid, giving a colorimetric endpoint that can be readspectrophotometrically。

Canine Insulin EIA 狗胰島素檢測(cè)試劑盒MATERIAL REQUIRED BUT NOT PROVIDED

• Pipettes with appropriate volumes (repeating pipettes preferred for addition ofenzyme conjugate 1X solution, Substrate TMB and Stop Solution)• Tubes, beakers and cylinders for reagent preparation• Redistilled water• Magnetic stirrer• Vortex mixer• Microplate reader with 450 nm filter• Microplate shaker (700–900 cycles per minute, orbital movement)• Microplate washing device with overflow function (recommended but not required)