Reactivity | HuSpecies Glossary |
Applications | Bioactivity |
Format | Carrier-Free |
Details of Functionality | Measured by the ability of the immobilized protein to support the adhesion of NIH‑3T3 mouse embryonic fibroblast cells. When 5 x 104 cells/well are added to recombinant human Nectin-4 coated plates (10 µg/mL with 100 µL/well), approximately >45% will adhere after 1 hour incubation at 37 °C. Optimal dilutions should be determined by each laboratory for each application. |
Source | Mouse myeloma cell line, NS0-derived human Nectin-4 protein Gly27-Val351, with a C-terminal 6-His tag |
Accession # | |
N-terminal Sequence | Gly27 |
Structure / Form | Monomer |
Protein/Peptide Type | Recombinant Proteins |
Gene | NECTIN4 |
Purity | >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. |
Endotoxin Note | <0.10 EU per 1 μg of the protein by the LAL method. |
Dilutions |
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Theoretical MW | 35.6 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
SDS-PAGE | 44 kDa, reducing conditions |
Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Buffer | Lyophilized from a 0.2 μm filtered solution in PBS. |
Purity | >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. |
Reconstitution Instructions | Reconstitute at 200 μg/mL in PBS. |
Nectin-4 (gene name PVRL4, poliovirus receptor-like 4) is a 66 kDa type I transmembrane glycoprotein belonging to the Nectin family of Ig superfamily proteins (1). The Latin word necto means “to connect”, indicating the role of nectins in Ca2+‑independent cell-cell adhesion (2). Nectin-4 forms homodimers in cis, followed by interactions in trans with Nectin-1 or -4 (1-3). Human Nectin-4 mRNA is normally expressed in the placenta, especially in endothelial cells, while in the mouse it is found in the embryo, lung, testis and brain (1, 4, 5). Human Nectin-4 cDNA encodes 510 amino acids (aa), including a 31 aa signal sequence, a 318 aa extracellular domain (ECD), a 21 aa transmembrane segment (TM), and a 140 aa cytoplasmic region. Nectin ECDs contain three Ig-like domains: an N‑terminal V-type that mediates ligand binding, and two C2-type (1, 3). One Nectin‑4 isoform lacks aa 412-436 in the cytoplasmic domain (1). In many human ductal breast or non-small cell lung carcinomas, Nectin‑4 is upregulated and a soluble 43 kDa form is found in the plasma (4-6). This form is generated from the membrane protein via the action of TACE/ADAM-17 (6). The extracellular domain of human Nectin-4 shares 91%, 92%, 93%, 91% and 90% amino acid sequence homology with the corresponding regions of mouse, rat, canine, porcine and bovine Nectin-4, respectively. In forming adherens junctions, trans interactions of Nectin-4 initiate cell-cell interactions and recruit intracellular cadherins through afadin and other junctional proteins (1, 2). These interactions organize the actin cytoskeleton, strengthen attachment to basement membrane and promote further cell-cell connections (2, 7). In humans, mutation of Nectin-4 has been correlated with ectodermal dysplasia-syndactyly syndrome, indicating a role for Nectin-4 in human development (7).
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