Human Nectin‑4 Antibody
R&D Systems | Catalog # AF2659
Key Product Details
Validated by
Species Reactivity
Validated:
Cited:
Applications
Validated:
Cited:
Label
Antibody Source
Product Specifications
Immunogen
Gly27-Val351
Accession # Q96NY8
Specificity
Clonality
Host
Isotype
Scientific Data Images for Human Nectin‑4 Antibody
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 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 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
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
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
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
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
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
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
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
CyTOF-ready
Flow Cytometry
Sample: MCF‑7 human breast cancer cell line
Immunohistochemistry
Sample: Immersion fixed paraffin-embedded sections of human placenta subjected to Antigen Retrieval Reagent-Basic (Catalog # CTS013)
Western Blot
Sample: Recombinant Human Nectin‑4 (Catalog # 2659-N4)
Flow Cytometry Panel Builder
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Advanced Features
- Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
- Spillover Popups - Visualize the spectra of individual fluorochromes
- Antigen Density Selector - Match fluorochrome brightness with antigen density
Formulation, Preparation, and Storage
Purification
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
Formulation
Shipping
Stability & Storage
- 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
Background: Nectin-4
Long Name
Alternate Names
Gene Symbol
UniProt
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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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- 7-Amino Actinomycin D (7-AAD) Cell Viability Flow Cytometry Protocol
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- Detection & Visualization of Antibody Binding
- Extracellular Membrane Flow Cytometry Protocol
- Flow Cytometry Protocol for Cell Surface Markers
- Flow Cytometry Protocol for Staining Membrane Associated Proteins
- Flow Cytometry Staining Protocols
- Flow Cytometry Troubleshooting Guide
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Intracellular Flow Cytometry Protocol Using Alcohol (Methanol)
- Intracellular Flow Cytometry Protocol Using Detergents
- Intracellular Nuclear Staining Flow Cytometry Protocol Using Detergents
- Intracellular Staining Flow Cytometry Protocol Using Alcohol Permeabilization
- Intracellular Staining Flow Cytometry Protocol Using Detergents to Permeabilize Cells
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Propidium Iodide Cell Viability Flow Cytometry Protocol
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Characterization of Human Th22 Cells
- Protocol for the Characterization of Human Th9 Cells
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- Protocol: Annexin V and PI Staining by Flow Cytometry
- Protocol: Annexin V and PI Staining for Apoptosis by Flow Cytometry
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Fluorokine Flow Cytometry Kits
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars