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|Method & Line||Sample & Target||Product||Package Info||Product||Package Info|
|IMMUNOgen - Immunoassays||Plasma,Serum||Insulin||Tests per Package: 100|
|IMMUNOgen||The Eclectica Insulin assay for the Eclectica and Eclectica TiCA analysers has been designed for the quantitative determination of Insulin in human serum and heparinized plasma. The method can be used to measure Insulin concentrations over the range of 2 to 200 µIU/ml. Primary calibration is against the WHO 1st International Reference Preparation 66/304 for Insulin.||Code: 1706200||Package: 2 x 50|
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The Eclectica Insulin assay for the Eclectica analyzers has been designed for the quantitative determination of Insulin in human serum and heparinized plasma. The method can be used to measure Insulin concentrations over the range of 2 to 200 µIU/ml. Primary calibration is against the WHO 1st International Reference Preparation 66/304 for Insulin.
Insulin is a polypeptide with a molecular weight of about 6000 Daltons. It is a heterodimer consisting of two different chains, the A-chain (21 residues) and the B-chain (30 residues), linked by disulphide bounds at position A7-B7 and A20-B19 (1, 2, 3, 4).
It is produced by the Langherans islets ß-cells of the Pancreas from its precursor molecule Proinsulin, which consists of the A and B chains linked by a connecting peptide known as C-Peptide. The Proinsulin is converted to Insulin by an enzymatic cleavage from the C-Peptide. It is stored in granules of the ß-cells, as hexamers, coordinated by the divalent metal ions Zn and Ca; as insulin leaves the cells and enters in the blood, dissociates in a monomeric form and circulates freely available for the binding to the receptors on the plasma membrane cells (5).
Insulin has many actions on body metabolism:
- it stimulates glycogenesis in muscle cells and liver;
- it stimulates liver glicolysis, so increases fatty acids synthesis;
- it facilitates glucose entry in muscle cells and adypocites;
- it facilitates ramified lateral chain aminoacids (valine, leucine, isoleucine) entry in muscle cells for the building of muscle proteins;
- it stimulates proteins synthesis.
Insulin assay can be considered a valid diagnostic tool for many dysmetabolic conditions:
- type I Diabetes (IDDM, Insulin-dependent-Diabetes-Mellitus), characterised by a lack of insulin production caused by the total destruction of the ß-cells: the insulin levels are very low (6).
- type II Diabetes (NIDDM, Non-Insulin-Dependent-Diabetes-Mellitus) in which insulin production is inadequate for the normal metabolic demand; in the early stage insulin levels may be normal or even increased, associated with insulin resistence, while in the over Diabetes, insulin levels are low (7).
- Insulinoma, a pancreatic endocrine tumour that secretes insulin autonomously. In this pathology, basal and after glucose administration, levels of insulin are higher than normal ranges (8, 9).
Increased insulin concentrations are also typical of clinical conditions such as insulin-treated IDMM, (ectopically) pancreas transplanted IDMM, obesity, stress and trauma (10, 11, 12, 14, 14).
- IGT (Impaired Glucose Tolerance), a condition in which the plasmatic levels of glucose, basal and after oral glucose stimulation, are between normal and diabetic values. It has been reported that 10% of patients with IGT become diabetic within 3 years and 30-50% become diabetic over their life (15).
The Eclectica assays for the Eclectica analyzers consists of:
In the Eclectica Insulin assay, two high affinity monoclonal antibodies are used in an immunoenzymetric assay system which incorporates magnetic solid phase separation. All incubation times and reagent volumes are determined by an assay specific software protocol. Fixed amounts of fluorescein-conjugated anti-Insulin monoclonal antibody and anti-Insulin monoclonal antibody conjugated to alkaline phosphatase are added to a sample, control, or standard. The reactants are incubated at 37°C. During the incubation fluorescein-anti-Insulin monoclonal antibody binds to a discrete site on the Insulin molecule. Alkaline phosphatase anti-Insulin monoclonal antibody binds to a second site on the Insulin molecule forming a sandwich.
At the end of the incubation period anti-fluorescein coupled to a magnetic solid phase is added in excess. This rapidly and specifically binds to the Insulin monoclonal antibody complex and is sedimented in a magnetic field.
After aspirating the liquid phase and washing the solid phase, a solution of the enzyme substrate, phenolphthalein monophosphate is added to the incubation cell and the each cell incubated at 37°C. After incubation the enzyme reaction is stopped by the addition of a stop reagent and the intensity of the color developed is measured photometrically. The intensity of the color developed is directly proportional, within the working range of the assay, to the concentration of Insulin in the sample. The concentration of Insulin in a patient sample or control is then determined by interpolation from a stored standard curve.
Eclectica Insulin Reagent Kit
Eclectica Insulin Calibration Kit
Eclectica Diluent 4 Kit
Eclectica Immunoassay Common Reagents Kit
Eclectica Immunoassay Wash Solution
Eclectica Systemic Wash Solution
1700006 Eclectica Cleaning Kit