Western blot analysis of extracts from various cell lines using Nectin-4/PVRL4 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, Nectin-4/PVRL4 expression is not detected in either MDA-MB-231 or A-549 cells.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human Nectin-4/PVRL4 protein (hNectin-4/PVRL4-Myc/DDK; +), using Nectin-4/PVRL4 Antibody (upper) and DYKDDDDK Tag Antibody #2368 (lower).
Western blot analysis of extracts from HT-1376 cells, untreated (-) or treated with PNGase F (+), using Nectin-4/PVRL4 Antibody.
|MW (kDa)||62, 75|
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Nectin-4/PVRL4 Antibody recognizes endogenous levels of total Nectin-4/PVRL4 protein. Based upon sequence alignment, this antibody is not predicted to cross-react with PVR, PVRL1, PVRL2, and PVRL3 proteins.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human Nectin-4/PVRL4 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Nectin-4/Poliovirus Receptor-Like 4 (PVRL4) is a type-I transmembrane glycoprotein that belongs to the immunoglobulin superfamily and promotes cell-cell adhesion by serving as a major component of adherens junctions (1-3). The extracellular domain of Nectin-4, which contains an Ig variable-like domain (V) and two Ig constant-like domains (C), mediates binding to the measles virus (4) and to neighboring cells through trans heterophilic interactions with Nectin-1 (5,6). Unlike other nectin family members, which are widely expressed in adult tissues, Nectin-4 expression in humans is largely restricted to the placenta (7). Research studies have demostrated that Nectin-4 is overexpressed in a variety of human solid tumors of the pancreas (8), breast (9,10), lung (11), and ovary (12). Due to its restricted expression pattern in normal human tissues, Nectin-4 may serve as a novel diagnostic and therapeutic target for a variety of human tumors.
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