The Calbiotech Insulin ELISA Kit is intended for the quantitative measurement Insulin in human serum or plasma.


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Product Catalog No: IS130D Pack Size: 96 Wells

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Summary

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. 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.

Test Principle

The 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 enzyme (HRP)-conjugated anti-insulin antibody and anti-insulin antibody bound to micro-titration well. A simple washing step removes unbound enzyme labeled antibody. The bound HRP complex is detected by reaction with TMB substrate. The reaction is stopped by adding acid to give a colorimetric endpoint that is read using ELISA reader.

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References
  1. Ashby, J. and Frier, B.: Circulating C-Peptide: Measurement and Clinical Applications. Annals of Clinical Biochemistry. 18:125, 1981
  2. Beischer, W.: Proinsulin and C-Peptide in Humans. Hormones in Normal and Abnormal Human Tissues. Volume 3K, Fotherby and Pal, S., ed. (Berlin: Walter DeGruyter). pp. 1-43, 1983
  3. Beyer, J., Krause V., Cordes V.: C-Peptide: Its Biogenesis, Structure, Determination and Clinical Significance. Giornale Italiano di Chimica Clinica 4 Supp. 9:22, 1979
  4. Bonger, A. and Garcia-Webb, P.: C-Peptide Measurement: Methods and Clinical Utility. CRC Critical Reviews in Clinical Laboratory Sciences. 19:297, 1984.
  5. Chevenne D., Ruiz J., Lohmann L., et.al.: Immunoradiometric Assay of Human Intact Proinsulin Applied to Patients with Type 2 Diabetes, Impaired Glucose Tolerance, and Hyperandrogenism. Clinical Chemistry. 40/5:754, 1994
  6. Bowsher R. R., Wolny J. D. and Frank B. H.: A Rapid and Sensitive Radioimmunoassay for the Measurement of Proinsulin in Human Serum. Diabetes. 41:1084, 1992
  7. Kao P. C., Taylor R. L. and Service F. J.: Proinsulin by Immunochemiluminometric Assay for the Diagnosis of Insulinoma. Jorunal of Clinical Endocrinology and Metabolism. 78:1048, 1994
  8. Dhahir F. J., Cook D. B. and Self C. H.: Amplified Enzyme-Linked Immunoassay of Human Proinsulin in Serum (Detection Limit: 0,1 pmol/L). Clinical Chemistry. 38/2:227, 1992
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