Description
Description: This assay employs a two-site sandwich ELISA to quantitate CANX in samples. An antibody specific for CANX has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CANX present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for CANX 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 CANX bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: Calnexin is a 90kDa integral protein of the endoplasmic reticulum. It consists of a large N-terminal calcium-binding lumenal domain, a single transmembrane helix and a short, acidic cytoplasmic tail. Calnexin is one of the chaperone molecules, which are characterized by their main function of assisting protein folding and quality control, ensuring that only properly folded and assembled proteins proceed further along the secretory pathway. Oligosaccharides with three sequential glucose residues are added to asparagine residues of the nascent proteins in the ER. The monoglucosylated oligosaccharides that are recognized by calnexin result from the trimming of two glucose residues by the sequential action of two glucosidases, I and II