Ultrasensitive Insulin ELISA
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INTENDED USE DRG
Ultrasensitive Insulin ELISA provides a method for the quantitative determination of insulin in human serum or plasma.
SUMMARY AND EXPLANATION OF THE TEST
Insulin is the principal hormone responsible for the control of glucose metabolism. It is synthesized in the ß-cells of the islets of Langerhans as the precursor, proinsulin, which is processed to form 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 B chain (21 and 30 amino acids respectively).The two chains are linked together by two inter-chain disulphide bridges.
There is also an intra-chain disulphide bridge in the A chain. Secretion of insulin is mainly controlled by plasma glucose concentration, and the hormone has a number of important metabolic actions. Its principal function is to control the uptake and utilisation of glucose in peripheral tissues via the glucose transporter. This and other hypoglycaemic activities, such as the inhibition of hepatic gluconeogenesis and glycogenolysis are counteracted by the hyperglycaemic hormones including glucagon, epinephrine (adrenaline), growth hormone and cortisol. Insulin concentrations are severely reduced in insulin-dependent diabetes mellitus (IDDM) and some other conditions such as hypopituitarism. Insulin levels are raised in non-insulin-dependent diabetes mellitus (NIDDM), obesity, insulinoma and some endocrine dysfunctions such as Cushing’s syndrome and acromegaly.
PRINCIPLE OF THE PROCEDURE DRG
Ultrasensitive Insulin ELISA is a solid phase two-site enzyme immunoassay. It is based on the direct sandwich technique in which two monoclonal antibodies are directed against separate antigenic determinants on the insulin molecule. During incubation insulin in the sample reacts with peroxidase-conjugated anti-insulin antibodies and antiinsulin antibodies bound to microplate. A simple washing step removes unbound enzyme labelled antibody. The bound conjugate is detected by reaction with 3,3′,5,5′-tetramethylbenzidine. The reaction is stopped by adding acid to give a colorimetric endpoint that is read spectrophotometrically.
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