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Flow cytometric analysis of cell-surface Nectin-4 on T47D (human ductal breast epithelial tumor epithelial) cells. T47D cells were stained with recombinant rabbit anti–Nectin-4 monoclonal antibody (NECTIN4/13438R) at dilutions of 1:50 (4 μg/ml; dark red), 1:100 (2 μg/ml; orange), 1:200 (1 μg/ml; light green) and 1:400 (0.5 μg/ml; dark green), then incubated with goat anti-rabbit IgG–CF488. CF488 fluorescence intensity (log₁₀ scale; x-axis) is plotted against relative cell count (y-axis). Progressive rightward shifts with increasing antibody concentration demonstrate dilution-dependent binding of Nectin-4. The blue histogram corresponds to the isotype control.
Homologous to the poliovirus receptor (PVR/CD155), the nectin immunoglobulin superfamily comprises four known isoforms (-1, -2, -3, and -4). The ectodomain of nectin family members comprises three Ig-like domains (V, C, C). Nectins localize at the adherens junctions (AJ) in epithelial and endothelial cells where they serve as adhesion molecules. Actin-based AJs play a role in mechanical adhesion, cellular morphogenesis and cellular differentiation. Nectin associates with the Actin cytoskeleton through its interaction with the Actin filament-binding protein afadin. Nectin 4 and afadin co-localize at cadherin-based adherens junctions in MDCKII epithelial cells. Nectin 4 and Nectin 3 share a common binding region in the V domain of Nectin 1 and thus compete for Nectin 1 binding. The Nectin 3/4 binding domain maps to the C-C’-C”-D b strands of the V domain of Nectin 1. Unlike other nectins, which are more widely expressed, Nectin 4 is mainly expressed in the placenta.
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