Description
Introduction: Ghrelin, a novel growth hormone releasing peptide is an acylated peptide that stimulates the release of growth hormone from the pituitary. It was isolated from rat stomach and the structure was determined as a peptide consisting of 28-amino acids. The Ser3 residue of Ghrelin is modified by n-octanoic acid, a modification necessary for hormone activity.
Principle of the Assay: The microtiter plate provided in this kit has been pre-coated with an antibody specific to Ghrelin. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for Ghrelin and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB (3,3',5,5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain Ghrelin, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm +/- 2 nm. The concentration of Ghrelin in the samples is then determined by comparing the O.D. of the samples to the standard curve