Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate WFS1 in samples. An antibody specific for WFS1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyWFS1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for WFS1 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 WFS1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Human WHDC1 expressed in insect cells had an apparent molecular mass of about 100 kD by SDS-PAGE. Western blot analysis detected WHDC1 in most human and mouse organs examined, with highest expression in brain. WHDC1 was also expressed in all cultured cell lines tested. WHDC1 partitioned predominantly in the membrane fraction of COS-7 cells, and it appeared to be a peripheral membrane protein, since high salt or alkaline pH released it into the soluble fraction. Immunohistochemical analysis localized WHDC1 primarily to the perinuclear compartment, near the microtubule-organizing center. In some cells, it also localized to tubulovesicular structures in the cell periphery. WHDC1 colocalized extensively with the cis-Golgi marker GM130 (GOLGA2), but not with medial- or trans-Golgi markers.
