Description
Principle of the assay: This assay employs a two-site sandwich ELISA to quantitative AG in Human serum, plasma. An antibody specific for AG has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any AG present is bound by the immobilized antibody. After removing any unbound substances, a biotin - conjugated antibody specific for AG is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of AG bound in the initial step. The color development is stopped and the intensity of the color is measured.
Background: Ghrelin is present in the peripheral circulation in two forms: acylated and non-acylated. The Acylated Ghrelin ELISA specifically measures the acylated form of ghrelin. The Acylated Ghrelin ELISA is based on a double-antibody sandwich technique; the Wells of the plate supplied with the kit are coated with a monoclonal antibody specific to the C-terminal part of ghrelin. This antibody will bind to any ghrelin introduced into the wells (standard or sample). The AChE - Fab` conjugate which recognizes the N-terminal part of acylated ghrelin is also added to the wells. This allows the two antibodies to form a sandwich by binding on different parts of the human acylated ghrelin. Each kit contains ghrelin (acylated) AChE-Fab` conjugate, human acylated ghrelin standard, human acylated ghrelin quality control, Ellman's reagent, EIA buffer, wash buffer, Tween 20, plates pre-coated with anti-ghrelin mouse monoclonal antibody, and complete instructions