Anti Insulin  

ELISA Results of Mab anti-Insulin antibody tested against human insulin by ELISA. Each well was coated with 0.1μg of conjugate. The starting concentration of antibody in the dilution series was 10 μg/ml. Each point on the Y-axis represents a 3-fold dilution. The midpoint of the titration curve represents approximately 5ng/ml antibody or a 1:200,000 dilution from the stock concentration. HRP conjugated Gt-a-Mouse IgG H&L(p/n 610-103-121) and TMB substrate were used for detection.

Dot blotting. Mab anti-Insulin antibody (clone 2D11.H5) is shown to detect human insulin by dot blot. Each dot blot represents 1 μl of non-denatured human insulin at various dilutions starting at 1.0 μg/ml spotted on to nitrocellulose. A 1:400 dilution of Mab anti-Insulin is used for 2 hour followed by detection using a 1:5,000 dilution of IRDyeTM800 conjugated Goat-a-Mouse IgG [H&L] (610-132-121) reacted for 45 min at room temperature and visualization using the OdysseyR Infrared Imaging System developed by LI-COR. Other detection systems will yield similar results. IRDye is a trademark of LI-COR, Inc.

Insulin is a polypeptide hormone produced by cells in the islets of Langerhans in the pancreas. Insulin decreases the levels of glucose in the blood and regulates the metabolism of glucose, fats and proteins. In order to meet the demand for insulin needed by diabetics, the hormone is mass-produced with the aid of genetically engineered bacteria, but can also be taken from pigs and cattle. The diagram to the left illustrates the structure of insulin and proinsulin. Proinsulin is a single polypeptide chain of 86 amino acids that permits correct alignment of three pairs of disulfide bonds. Insulin is derived from proinsulin by cleavage of the C-peptide structure at the dipeptides Arg-Arg and Lys-Arg. Insulin is composed of an A chain of 21 amino acids and a B chain of 30 amino acids, the chains being held together by two disulfide bonds. A third disulfide bond is present within the A chain.