Human Nectin-4 Antibody

R&D Systems | Catalog # AF2659

R&D Systems

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Porcine, Canine, Hamster, Primate - Chlorocebus aethiops (African Green Monkey), Primate - Macaca fascicularis (Crab-eating Monkey or Cynomolgus Macaque), Xenograft

Applications

Validated:

Immunohistochemistry, Western Blot, Flow Cytometry, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Functional Assay, Westen Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Nectin-4 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Nectin-4
Gly27-Val351
Accession # Q96NY8

Specificity

Detects human Nectin-4 in direct ELISAs and Western blots. In direct ELISAs, approximately 75% cross‑reactivity with recombinant mouse Nectin-4 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human Nectin-4 Antibody

Nectin‑4 antibody in Human Placenta by Immunohistochemistry (IHC-P).

Nectin‑4 in Human Placenta.

Nectin-4 was detected in immersion fixed paraffin-embedded sections of human placenta using 10 µg/mL Goat Anti-Human Nectin-4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2659) overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Nectin-4 in MCF-7 Human Cell Line by Flow Cytometry.

MCF-7 human breast adenocarcinoma cell line was stained with Goat Anti-Human Nectin-4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2659, filled histogram) or control antibody (AB-108-C, open histogram), followed by PE-conjugated Anti-Goat IgG Secondary Antibody (F0107).
Detection of Human Nectin-4/PVRL4 by Western Blot

Detection of Human Nectin-4/PVRL4 by Western Blot

Inhibition of Nectin-4 expression by gene silencing decreases cell proliferation in pancreatic cancer cells. (A,B) Capan-2 and BxPC-3 cells were transfected with Nectin-4 siRNA or control RNA. The relative expression of Nectin-4 was significantly reduced in both cells when transfected with Nectin-4 siRNA for up to 72 hours as determined by quantitative real-time PCR and Western blot analysis (n = 4 of each group). (C) Cell proliferation was significantly inhibited by Nectin-4 gene silencing in both cells after 72 hours incubation as determined by MTS assay (n = 6 of each group). *P < 0.001 versus control siRNA (Student’s t test). Image collected and cropped by CiteAb from the following publication (https://jeccr.biomedcentral.com/articles/10.1186/s13046-015-0144-7), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Nectin-4/PVRL4 by Western Blot

Detection of Human Nectin-4/PVRL4 by Western Blot

Knockdown expression and over-expression of Nectin-4 in human EC cell lines Eca-109 and TE-1. a The knockdown expression of Nectin-4 at mRNA level by using RNAi approach in human EC lines was confirmed by real-time RT-PCR, and the Nectin-4 mRNA expression level in LV-Nectin-4-shRNA group cells was significantly lower than that in LV-NC group cells (both in Eca-109 and in TE-1, P < 0.0001). b The decreased Nectin-4 protein expression after knockdown in human EC cell lines Eca-109 and TE-1, were confirmed by using western blot method. c The Nectin-4 protein expression level in LV-Nectin-4-shRNA group cells was significantly lower than that in LV-NC group cells (in Eca-109, P < 0.01, and in TE-1, P < 0.001). d The over-expression of Nectin-4 at mRNA level in human EC lines was confirmed by real-time RT-PCR, which showed that the increased Nectin-4 mRNA expression level in LV-Nectin-4-OE group cells compared with LV-Vector-Ctrl group cells (both in Eca-109 and in TE-1, P < 0.0001). e The increased Nectin-4 protein expression in human EC cell lines Eca-109 and TE-1 were confirmed by using western blot method. f The Nectin-4 protein expression level in LV-Nectin-4-OE group cells was significantly higher than that in LV-Vector-Ctrl group cells (in Eca-109, P < 0.01, and in TE-1, P < 0.001) Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31043861), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Nectin-4/PVRL4 by Immunohistochemistry

Detection of Human Nectin-4/PVRL4 by Immunohistochemistry

Nectin-4 expression in EC tissues. a, b Positive staining of Nectin-4 could be found in the cytoplasm and on the membrane of cancer cells, while weak or negative staining of Nectin-4 was found in normal esophageal tissues. c The staining intensity of Nectin-4 in EC tissues was significantly higher than that in adjacent normal tissues (P < 0.0001). d The survival analysis showed that the overall survival rate of the patients with higher Nectin-4 expression was significantly poorer compared with those showing lower Nectin-4 expression (HR = 1.704, 95% CI 1.027–2.825, P = 0.039) Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31043861), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Nectin-4/PVRL4 by Immunohistochemistry

Detection of Human Nectin-4/PVRL4 by Immunohistochemistry

Nectin-4 expression in EC tissues. a, b Positive staining of Nectin-4 could be found in the cytoplasm and on the membrane of cancer cells, while weak or negative staining of Nectin-4 was found in normal esophageal tissues. c The staining intensity of Nectin-4 in EC tissues was significantly higher than that in adjacent normal tissues (P < 0.0001). d The survival analysis showed that the overall survival rate of the patients with higher Nectin-4 expression was significantly poorer compared with those showing lower Nectin-4 expression (HR = 1.704, 95% CI 1.027–2.825, P = 0.039) Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31043861), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Nectin-4/PVRL4 by Immunocytochemistry/Immunofluorescence

Detection of Human Nectin-4/PVRL4 by Immunocytochemistry/Immunofluorescence

Nectin-4 expression in human pancreatic cancer tissues. (A,B) Tumor cell with high or low Nectin-4 expression. (C) There was limited expression of Nectin-4 in non-cancer tissues including islet cells, acinar cells and normal gland. Representative case of tumor Nectin-4 expression. Original magnification, ×200. Image collected and cropped by CiteAb from the following publication (https://jeccr.biomedcentral.com/articles/10.1186/s13046-015-0144-7), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Nectin-4/PVRL4 by Immunocytochemistry/Immunofluorescence

Detection of Human Nectin-4/PVRL4 by Immunocytochemistry/Immunofluorescence

Nectin-4 expression in human pancreatic cancer tissues. (A,B) Tumor cell with high or low Nectin-4 expression. (C) There was limited expression of Nectin-4 in non-cancer tissues including islet cells, acinar cells and normal gland. Representative case of tumor Nectin-4 expression. Original magnification, ×200. Image collected and cropped by CiteAb from the following publication (https://jeccr.biomedcentral.com/articles/10.1186/s13046-015-0144-7), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Nectin-4/PVRL4 by Western Blot

Detection of Human Nectin-4/PVRL4 by Western Blot

Knockdown expression and over-expression of Nectin-4 in human EC cell lines Eca-109 and TE-1. a The knockdown expression of Nectin-4 at mRNA level by using RNAi approach in human EC lines was confirmed by real-time RT-PCR, and the Nectin-4 mRNA expression level in LV-Nectin-4-shRNA group cells was significantly lower than that in LV-NC group cells (both in Eca-109 and in TE-1, P < 0.0001). b The decreased Nectin-4 protein expression after knockdown in human EC cell lines Eca-109 and TE-1, were confirmed by using western blot method. c The Nectin-4 protein expression level in LV-Nectin-4-shRNA group cells was significantly lower than that in LV-NC group cells (in Eca-109, P < 0.01, and in TE-1, P < 0.001). d The over-expression of Nectin-4 at mRNA level in human EC lines was confirmed by real-time RT-PCR, which showed that the increased Nectin-4 mRNA expression level in LV-Nectin-4-OE group cells compared with LV-Vector-Ctrl group cells (both in Eca-109 and in TE-1, P < 0.0001). e The increased Nectin-4 protein expression in human EC cell lines Eca-109 and TE-1 were confirmed by using western blot method. f The Nectin-4 protein expression level in LV-Nectin-4-OE group cells was significantly higher than that in LV-Vector-Ctrl group cells (in Eca-109, P < 0.01, and in TE-1, P < 0.001) Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31043861), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Nectin-4/PVRL4 by Immunocytochemistry/Immunofluorescence

Detection of Human Nectin-4/PVRL4 by Immunocytochemistry/Immunofluorescence

Nectin-4 expression in human pancreatic cancer tissues. (A,B) Tumor cell with high or low Nectin-4 expression. (C) There was limited expression of Nectin-4 in non-cancer tissues including islet cells, acinar cells and normal gland. Representative case of tumor Nectin-4 expression. Original magnification, ×200. Image collected and cropped by CiteAb from the following publication (https://jeccr.biomedcentral.com/articles/10.1186/s13046-015-0144-7), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Nectin-4 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: MCF‑7 human breast cancer cell line

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human placenta subjected to Antigen Retrieval Reagent-Basic (Catalog # CTS013)

Western Blot

0.1 µg/mL
Sample: Recombinant Human Nectin‑4 (Catalog # 2659-N4)

Flow Cytometry Panel Builder

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Advanced Features

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Nectin-4

Nectin-4 is a type I transmembrane glycoprotein belonging to the Nectin family of Ig superfamily proteins. It is both a homophilic and heterophilic (with Nectin-1) cell adhesion molecule that is expressed in the embryo and in breast carcinoma. A soluble form of Nectin-4 is generated from the membrane protein via the action of TACE/ADAM-17. The extracellular domain of human Nectin-4 shares 90% and 92% amino acid sequence homology with the corresponding regions of mouse and rat Nectin-4, respectively.

Long Name

Poliovirus Receptor Related 4

Alternate Names

LNIR, Nectin4, PRR4, PVRL4

Entrez Gene IDs

81607 (Human); 71740 (Mouse); 498281 (Rat); 102127936 (Cynomolgus Monkey)

Gene Symbol

NECTIN4

UniProt

Q96NY8

Additional Nectin-4 Products

Product Documents for Human Nectin-4 Antibody

Certificate of Analysis

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Product Specific Notices for Human Nectin-4 Antibody

For research use only

Citations for Human Nectin-4 Antibody

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