Nectin-4/PVRL4 Antibody [Unconjugated]

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.

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).

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 (//jeccr.biomedcentral.com/articles/10.1186/s13046-015-0144-7), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/31043861), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/31043861), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/31043861), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//jeccr.biomedcentral.com/articles/10.1186/s13046-015-0144-7), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//jeccr.biomedcentral.com/articles/10.1186/s13046-015-0144-7), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/31043861), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//jeccr.biomedcentral.com/articles/10.1186/s13046-015-0144-7), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, Flow, IHC, CyTOF-ready
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Nectin-4/PVRL4 Antibody [Unconjugated] Summary

• 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.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: PVRL4
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunohistochemistry 5-15 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Nectin-4/PVRL4 Antibody [Unconjugated]

• EDSS1
• Ig superfamily receptor LNIR
• LNIR
• LNIRnectin 4
• Nectin4
• Nectin-4
• poliovirus receptor-related 4
• poliovirus receptor-related protein 4
• PRR4
• PRR4DKFZp686K05193
• PVRL4

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Nectin-4/PVRL4 Antibody (AF2659)(23)

Publications using AF2659

• Tamura, K;Fujiyuki, T;Moritoh, K;Akimoto, H;Iizuka, K;Sato, H;Asano, K;Yoneda, M;Kai, C; Anti-tumor activity of a recombinant measles virus against canine lung cancer cells Scientific reports 2023-10-24 [PMID: 37875555] (Flow Cytometry, Xenograft)
• Kanako Moritoh, Koichiro Shoji, Yosuke Amagai, Tomoko Fujiyuki, Hiroki Sato, Misako Yoneda et al. Immune response elicited in the tumor microenvironment upon rMV‐SLAMblind cancer virotherapy Cancer Science 5/1/2023 [PMID: 36715555]
• V Gradauskai, M Inglebert, J Doench, M Scherer, M Dettwiler, M Wyss, N Shrestha, S Rottenberg, P Plattet LRP6 Is a Functional Receptor for Attenuated Canine Distemper Virus MBio, 2023-01-16;0(0):e0311422. 2023-01-16 [PMID: 36645301] (ICC, Chlorocebus aethiops (green monkey))
• Fanfan Li, Shuping Zhao, Cheng Wei, Yaodi Hu, Tianlong Xu, Xueyi Xin et al. Development of Nectin4/FAP-targeted CAR-T cells secreting IL-7, CCL19, and IL-12 for malignant solid tumors Frontiers in Immunology 11/21/2022 [PMID: 36479116]
• Punit Upadhyaya, Johanna Lahdenranta, Kristen Hurov, Sailaja Battula, Rachel Dods, Eric Haines et al. Anticancer immunity induced by a synthetic tumor-targeted CD137 agonist Journal for ImmunoTherapy of Cancer 1/26/2021 [PMID: 33500260]
• K Iizuka, K Shoji, T Fujiyuki, K Moritoh, K Tamura, A Yoshida, H Sato, M Yoneda, K Asano, C Kai Antitumor activity of an oncolytic measles virus against canine urinary bladder transitional cell carcinoma cells Research in Veterinary Science, 2020-10-22;0(0):. 2020-10-22 [PMID: 33183781] (IHC, Canine)
• H Deng, H Shi, L Chen, Y Zhou, J Jiang Over-expression of Nectin-4 promotes progression of esophageal cancer and correlates with poor prognosis of the patients Cancer Cell Int., 2019-04-23;19(0):106. 2019-04-23 [PMID: 31043861] (IHC, Human)
• Leonardo Della Salda, Marcella Massimini, Mariarita Romanucci, Chiara Palmieri, Antonella Perillo, Valeria Grieco et al. Nectin‐4 and p63 immunohistochemical expression in canine prostate tumourigenesis Veterinary and Comparative Oncology 9/1/2019 [PMID: 30767361]
• H Shiotani, M Miyata, Y Itoh, S Wang, A Kaito, A Mizoguchi, M Yamasaki, M Watanabe, K Mandai, H Mochizuki, Y Takai Localization of nectin-2? at the boundary between the adjacent somata of the clustered cholinergic neurons and its regulatory role in the subcellular localization of the voltage-gated A-type K+channel Kv4.2 in the medial habenula J. Comp. Neurol., 2018-03-30;0(0):. 3/30/2018 [PMID: 29524214]
• Blake A. Jacobson, Ahad A. Sadiq, Shaogeng Tang, Joe Jay-Dixon, Manish R. Patel, Jeremy Drees et al. Cap-dependent translational control of oncolytic measles virus infection in malignant mesothelioma Oncotarget 9/8/2017 [PMID: 28968974]
• W Pratakpiri, AP Ping Teh, A Radtanakat, N Pirarat, N Thi Lan, M Takeda, S Techangams, R Yamaguchi Expression of canine distemper virus receptor nectin-4 in the central nervous system of dogs Sci Rep, 2017-03-23;7(1):349. 2017-03-23 [PMID: 28336928] (IHC-P, Canine)
• M Kitayama, K Mizutani, M Maruoka, K Mandai, S Sakakibara, Y Ueda, T Komori, Y Shimono, Y Takai A Novel Nectin-mediated Cell Adhesion Apparatus That Is Implicated in Prolactin Receptor Signaling for Mammary Gland Development J. Biol. Chem, 2016-01-12;291(11):5817-31. 2016-01-12 [PMID: 26757815] (ICC, Mouse)
• Brajesh K. Singh, Andrew L. Hornick, Sateesh Krishnamurthy, Anna C. Locke, Crystal A. Mendoza, Mathieu Mateo et al. The Nectin-4/Afadin Protein Complex and Intercellular Membrane Pores Contribute to Rapid Spread of Measles Virus in Primary Human Airway Epithelia Journal of Virology 7/15/2015 [PMID: 25926640]
• Nishiwada S, Sho M, Yasuda S, Shimada K, Yamato I, Akahori T, Kinoshita S, Nagai M, Konishi N, Nakajima Y Nectin-4 expression contributes to tumor proliferation, angiogenesis and patient prognosis in human pancreatic cancer. J Exp Clin Cancer Res, 2015-03-28;34(0):30. 2015-03-28 [PMID: 25888293] (IHC-P, Human)
• Alves L, Khosravi M, Avila M, Ader-Ebert N, Bringolf F, Zurbriggen A, Vandevelde M, Plattet P SLAM- and Nectin-4-Independent Noncytolytic Spread of Canine Distemper Virus in Astrocytes. J Virol, 2015-03-18;89(10):5724-33. 2015-03-18 [PMID: 25787275] (IHC, Canine)
• Galanis E, Atherton P, Maurer M, Knutson K, Dowdy S, Cliby W, Haluska P, Long H, Oberg A, Aderca I, Block M, Bakkum-Gamez J, Federspiel M, Russell S, Kalli K, Keeney G, Peng K, Hartmann L Oncolytic measles virus expressing the sodium iodide symporter to treat drug-resistant ovarian cancer. Cancer Res, 2014-11-14;75(1):22-30. 2014-11-14 [PMID: 25398436] (IHC, Human)
• R Lattanzio, R Ghasemi, F Brancati, R L Sorda, N Tinari, L Perracchio et al. Membranous Nectin-4 expression is a risk factor for distant relapse of T1-T2, N0 luminal-A early breast cancer Oncogenesis 9/1/2014 [PMID: 25178039]
• Manish R. Patel, Blake A. Jacobson, Holly Belgum, Ahmad Raza, Ahad Sadiq, Jeremy Drees et al. Measles Vaccine Strains for Virotherapy of Non–Small-Cell Lung Carcinoma Journal of Thoracic Oncology 8/27/2014 [PMID: 25157763]
• Manish R. Patel, Blake A. Jacobson, Holly Belgum, Ahmad Raza, Ahad Sadiq, Jeremy Drees et al. Measles Vaccine Strains for Virotherapy of Non–Small-Cell Lung Carcinoma Journal of Thoracic Oncology 2014-08-27 [PMID: 25157763] (Neutralization, Human)
• Fortugno P, Josselin E, Tsiakas K, Agolini E, Cestra G, Teson M, Santer R, Castiglia D, Novelli G, Dallapiccola B, Kurth I, Lopez M, Zambruno G, Brancati F Nectin-4 mutations causing ectodermal dysplasia with syndactyly perturb the rac1 pathway and the kinetics of adherens junction formation. J Invest Dermatol, 2014-02-27;134(8):2146-53. 2014-02-27 [PMID: 24577405] (ICC, IHC, Human)
• Pavlova N, Pallasch C, Elia A, Braun C, Westbrook T, Hemann M, Elledge S A role for PVRL4-driven cell-cell interactions in tumorigenesis. Elife, 2013-04-30;2(0):e00358. 2013-04-30 [PMID: 23682311] (Functional Assay, IHC, Western Blot, Human, Mouse)
• Lethal canine distemper virus outbreak in cynomolgus monkeys in Japan in 2008. J Virol, 2012-11-07;87(2):1105-14. 2012-11-07 [PMID: 23135729] (IHC-P, Primate - Macaca fascicularis (Crab-eating Monkey or Cynomolgus Macaque))
• Noyce RS, Bondre DG, Ha MN Tumor cell marker PVRL4 (nectin 4) is an epithelial cell receptor for measles virus. PLoS Pathog., 2011-08-25;7(8):e1002240. 2011-08-25 [PMID: 21901103] (ICC, Neutralization, Western Blot, Hamster, Human)

Bioinformatics

• Gene Symbol: PVRL4
• Uniprot:
  • Q96NY8()