Human Nectin-1, NECTIN1 ELISA Kit

Poliovirus receptor-related 1 (herpesvirus entry mediator C; nectin-1; CD111), also known as PVRL1 is a cell adhesion molecule belonging to the immunoglobulin superfamily that can bind to virion glycoprotein D (gD) to mediate entry of herpes simplex viruses (HSV) and pseudorabies virus (PRV) . CD111/Nectin-1/PVRL1 colocalizes with E-cadherin at adherens junctions in epithelial cells. The disruption of cell junctions can result in the redistribution of nectin-1. To determine whether disruption of junctions by calcium depletion influenced the susceptibility of epithelial cells to viral entry, Madin-Darby canine kidney cells expressing endogenous nectin-1 or transfected human nectin-1 were tested for the ability to bind soluble forms of viral gD and to be infected by HSV and PRV, before and after calcium depletion. It has been revealed that binding of HSV and PRV gD was localized to adherens junctions in cells maintained in normal medium but was distributed, along with nectin-1, over the entire cell surface after calcium depletion. Both the binding of gD and the fraction of cells that could be infected by HSV-1 and PRV were enhanced by calcium depletion. Taken together, CD111/Nectin-1/PVRL1 confined to adherens junctions in epithelial cells is not very accessible to virus, whereas dissociation of cell junctions releases nectin-1 to serve more efficiently as an entry recptor.

Concentration | O.D. | Average | Corrected ------------------------------------------ 5000 pg/ml | 2.567 | 2.447 | 2.341 2.327 2500 pg/ml | 1.272 | 1.327 | 1.221 1.382 1250 pg/ml | 0.701 | 0.715 | 0.609 0.729 625 pg/ml | 0.431 | 0.408 | 0.302 0.384 312.5 pg/ml | 0.246 | 0.249 | 0.143 0.252 156.25 pg/ml | 0.197 | 0.187 | 0.081 0.177 78.125 pg/ml | 0.139 | 0.138 | 0.032 0.137 0 pg/ml | 0.1 | 0.106 | 0 0.112

Intra/Inter-Assay | Sample | n | Mean | Standard Deviation | CV% ---------------------------------------------------------------- Intra-assay | 1 | 20 | 523 | 37 | 7.1 Intra-assay | 2 | 20 | 1494 | 94.9 | 6.4 Intra-assay | 3 | 20 | 3735 | 226 | 6.1 Inter-assay | 1 | 20 | 528 | 33.3 | 6.3 Inter-assay | 2 | 20 | 1499 | 81.9 | 5.5 Inter-assay | 3 | 20 | 3779 | 261 | 6.9

Components | Quantity --------------------- Pre-coated ELISA Plate | 12 x 8 well strips x 1 Standard Solution | 0.2mL x 1 vail Dilution Buffer | 20mL x 1 vail Stop Solution | 6mL x 1 vail Substrate Solution A | 12mL x 1 vail Substrate Solution B | 12mL x 1 vail Wash Buffer Concentrate (25x) | 20mL x 1 vail Detection Antibody Concentrate | 0.6mL x 1 vail