Description
This assay employs a two-site sandwich ELISA to quantitate CGN in samples. An antibody specific for CGN has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CGN present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for CGN 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 CGN bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: Cingulin is a homodimer, each subunit containing a globular "head" domain, a long coiled-coil "rod" domain, and a small globular "tail". This domain organization is conserved throughout vertebrate species.However, cingulin homologs have not been detected in invertebrate species.Cingulin is specifically localized at tight junctions in epithelial cells, unlike ZO-1, which is also detected at adherens-type junctions in non-epithelial cells. During early Xenopus embryo development, cingulin is detected at a cortical localization, and then accumulates at apical junctions, unlike ZO-1 and other junctional proteins, that are targeted to the new regions of cell-cell contact via the lateral domain