Human/Mouse/Rat N-Cadherin Antibody

Catalog # Availability Size / Price Qty
AF6426
AF6426-SP
Detection of Human, Mouse, and Rat N‑Cadherin by Western Blot.
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Product Details
Citations (23)
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Human/Mouse/Rat N-Cadherin Antibody Summary

Species Reactivity
Human, Mouse, Rat
Specificity
Detects human, mouse, and rat N‑Cadherin in Western blots. In direct ELISA, less than 10% cross-reactivity with recombinant human
(rh) E-Cadherin, and rhR-Cadherin is observed.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human N‑Cadherin
Asp160-Ala724
Accession # P19022
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.5 µg/mL
See below
ELISA

This antibody functions as an ELISA detection antibody to detect human N-Cadherin when paired with Mouse Anti-Human N‑Cadherin Monoclonal Antibody (Catalog # MAB13883).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human N-Cadherin DuoSet ELISA Kit (Catalog # DY1388-05) for convenient development of a sandwich ELISA.

 
Flow Cytometry
0.25 µg/106 cells
A172 human glioblastoma cell line
Immunohistochemistry
1-15 µg/mL
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Immunocytochemistry
5-15 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human, Mouse, and Rat N-Cadherin antibody by Western Blot. View Larger

Detection of Human, Mouse, and Rat N‑Cadherin by Western Blot. Western blot shows lysates of A549 human lung carcinoma cell line, HeLa human cervical epithelial carcinoma cell line, C2C12 mouse myoblast cell line, and rat brain tissue. PVDF Membrane was probed with 0.5 µg/mL of Sheep Anti-Human/Mouse/Rat N-Cadherin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6426) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for N-Cadherin at approximately 130 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry N-Cadherin antibody in A549 Human Cell Line by Immunocytochemistry (ICC). View Larger

N‑Cadherin in A549 Human Cell Line. N-Cadherin was detected in immersion fixed A549 human lung carcinoma cell line using Sheep Anti-Human/Mouse/Rat N-Cadherin Affinity-purified Polyclonal Antibody (Catalog # AF6426) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry View Larger

N‑Cadherin in Human Heart N-Cadherin was detected in immersion fixed paraffin-embedded sections of human heart using Sheep Anti-Human/Mouse/Rat N-Cadherin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6426) at 1 µg/mL overnight at 4 °C. Tissue was stained using the Sheep IgG VisUCyte HRP Polymer Antibody (Catalog # VC006). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to intercalated disks between the myocardial cells. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Human N‑Cadherin ELISA Standard Curve. Recombinant Human N-Cadherin protein was serially diluted 2-fold and captured by Mouse Anti-Human N-Cadherin Monoclonal Antibody (Catalog # MAB13883) coated on a Clear Polystyrene Microplate (Catalog # DY990). Sheep Anti-Human/Mouse/Rat N-Cadherin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6426) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).

Flow Cytometry View Larger

Detection of N‑Cadherin in A172 cells by Flow Cytometry. A172 cells were stained with Sheep Anti-Human/Mouse/Rat N‑Cadherin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6426, filled histogram) or isotype control antibody (Catalog # 5-001-A, open histogram), followed by Fluorescein-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0125). View our protocol for Staining Membrane-associated Proteins.

Western Blot Detection of Human N-Cadherin by Western Blot View Larger

Detection of Human N-Cadherin by Western Blot N-Cadherin Depletion Rescues the Migration Defect in Rab14- and FAM116-Depleted Cells(A and B) A549 cells were treated with control, Rab14, FAM116A, N-cadherin, E-cadherin, and FGFR2 siRNA duplexes alone or in the combinations indicated for 72 hr (A). The cells were then western blotted for N-cadherin, E-cadherin, or tubulin as a loading control or (B) fixed and then stained with DAPI and antibodies to N-cadherin. Scale bar indicates 10 μm.(C) A549 cells were treated with control, Rab14, FAM116A, N-cadherin, E-cadherin, and FGFR2 siRNA duplexes alone or in the combinations indicated for 72 hr. The cell monolayers were then scratched and imaged for 16 hr. Cell migration was measured and is plotted on the graph with error bars to show the standard error of the mean (n = 3).(D) Images are shown from the 0 and 16 hr time points for the Rab14- and FAM116A-depleted cells in the presence and absence of N-cadherin siRNA. Scale bar indicates 50 μm.See also Figure S5. Image collected and cropped by CiteAb from the following publication (https://linkinghub.elsevier.com/retrieve/pii/S1534580712001876), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Human N-Cadherin by Western Blot View Larger

Detection of Human N-Cadherin by Western Blot Abnormal Adherens Junctions in Rab14- and FAM116-Depleted Cells(A) A549 cells were treated with control, Rab4, Rab11, Rab14, FAM116A, and Avl9 siRNA duplexes for 72 hr. The cells were lysed in SDS-PAGE sample buffer and then western blotted as indicated in the figure. Tubulin was used as a loading control.(B) A549 cells were treated with control, Rab14, and FAM116A siRNA duplexes for 72 hr, fixed, and then stained with DAPI and antibodies to N-cadherin.(C) A549 cells were treated with control, Rab4, Rab11, Rab14, FAM116A, and Avl9 siRNA duplexes for 72 hr. The cell monolayers were then scratched, samples fixed at 0, 4, 8, and 16 hr, and then stained with DAPI and antibodies to N-cadherin. Images are oriented so the wound is to the right. Scale bar indicates 10 μm in all images.See also Figure S6. Image collected and cropped by CiteAb from the following publication (https://linkinghub.elsevier.com/retrieve/pii/S1534580712001876), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.

Background: N-Cadherin

N-Cadherin (Neural Cadherin; also CD325 and Cadherin-2) is a 130-135 kDa member of the "classical" (or type I) cadherin subfamily, cadherin superfamily of proteins. It is expressed on multiple cell types, including neurons, fibroblasts, Schwann cells, endothelial cells and hepatic stellate cells. N-Cadherin mediates homotypic binding, either in cis (same cell) or trans (adjacent cell). proN-Cadherin is expressed as an 881 amino acid (aa) type I transmembrane glycoprotein. It may be initially inserted into the ER, where the 15-20 kDa prodomain (aa 26-159) is cleaved by proprotein convertase, and the mature molecule (aa 160-906) is transported to the surface. Mature N-Cadherin contains a 565 aa extracellular region (aa 160-724) that possesses five cadherin domains (aa 160-714), and a 161 aa cytoplasmic tail that undergoes phosphorylation at Tyr785. There is one splice variant that contains a 10 aa substitution for aa 839-906. Over aa 160-724, human N Cadherin shares 98% aa identity with mouse N-Cadherin.

Long Name
Neural Cadherin
Entrez Gene IDs
1000 (Human); 12558 (Mouse); 83501 (Rat)
Alternate Names
ACOGS; ARVD14; cadherin 2, type 1, N-cadherin (neuronal); Cadherin-2; calcium-dependent adhesion protein, neuronal; CD325 antigen; CD325; CDH2; CDHN; CDw325; NCAD; N-cadherin 1; NCadherin; N-Cadherin; Neural cadherin; neural-cadherin

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Citations for Human/Mouse/Rat N-Cadherin Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

23 Citations: Showing 1 - 10
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  1. The Wound Microenvironment Reprograms Schwann Cells to Invasive Mesenchymal-like Cells to Drive Peripheral Nerve Regeneration.
    Authors: Clements MP, Byrne E, Camarillo Guerrero LF et al.
    Neuron.
  2. GSK3 regulates hair cell fate in the developing mammalian cochlea
    Authors: Kathryn Ellis, Elizabeth C. Driver, Takayuki Okano, Abigail Lemons, Matthew W. Kelley
    Developmental Biology
  3. Cardiomyocyte FGF signaling is required for Cx43 phosphorylation and cardiac gap junction maintenance
    Authors: Takashi Sakurai, Mariko Tsuchida, Paul D. Lampe, Masahiro Murakami
    Experimental Cell Research
  4. Induction of in vitro Metabolic Zonation in Primary Hepatocytes Requires Both Near-Physiological Oxygen Concentration and Flux
    Authors: Benedikt Scheidecker, Marie Shinohara, Masahiro Sugimoto, Mathieu Danoy, Masaki Nishikawa, Yasuyuki Sakai
    Frontiers in Bioengineering and Biotechnology
  5. Synapse elimination activates a coordinated homeostatic presynaptic response in an autaptic circuit
    Authors: Cecilia D. Velasco, Artur Llobet
    Communications Biology
  6. Rab14 and Its Exchange Factor FAM116 Link Endocytic Recycling and Adherens Junction Stability in Migrating Cells
    Authors: Andrea Linford, Shin-ichiro Yoshimura, Ricardo Nunes Nunes Bastos, Lars Langemeyer, Andreas Gerondopoulos, Daniel J. Rigden et al.
    Developmental Cell
  7. Ym1+ macrophages orchestrate fibrosis, lesion growth and progression during development of murine pancreatic cancer
    Authors: Fleming Martinez A, DOppler H, Bastea L et al.
    iScience
  8. An integrated organoid omics map extends modeling potential of kidney disease
    Authors: Lassé, M;El Saghir, J;Berthier, CC;Eddy, S;Fischer, M;Laufer, SD;Kylies, D;Hutzfeldt, A;Bonin, LL;Dumoulin, B;Menon, R;Vega-Warner, V;Eichinger, F;Alakwaa, F;Fermin, D;Billing, AM;Minakawa, A;McCown, PJ;Rose, MP;Godfrey, B;Meister, E;Wiech, T;Noriega, M;Chrysopoulou, M;Brandts, P;Ju, W;Reinhard, L;Hoxha, E;Grahammer, F;Lindenmeyer, MT;Huber, TB;Schlüter, H;Thiel, S;Mariani, LH;Puelles, VG;Braun, F;Kretzler, M;Demir, F;Harder, JL;Rinschen, MM;
    Nature communications
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC/IF
  9. Integrating Oxygen and 3D Cell Culture System: A Simple Tool to Elucidate the Cell Fate Decision of hiPSCs
    Authors: RR Khadim, RK Vadivelu, T Utami, FG Torizal, M Nishikawa, Y Sakai
    International Journal of Molecular Sciences, 2022-06-30;23(13):.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  10. Specific Binding of Novel SPION-Based System Bearing Anti-N-Cadherin Antibodies to Prostate Tumor Cells
    Authors: K Karnas, T Str?czek, C Kapusta, M Lekka, J Duli?ska-L, A Karewicz
    International Journal of Nanomedicine, 2021-09-24;16(0):6537-6552.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  11. Reciprocal interplay between asporin and decorin: Implications in gastric cancer prognosis
    Authors: D Basak, Z Jamal, A Ghosh, PK Mondal, P Dey Talukd, S Ghosh, B Ghosh Roy, R Ghosh, A Halder, A Chowdhury, GK Dhali, BK Chattopadh, ML Saha, A Basu, S Roy, C Mukherjee, NK Biswas, U Chatterji, S Datta
    PLoS ONE, 2021-08-11;16(8):e0255915.
    Species: Human
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  12. Inhibition of Sema4D/PlexinB1 signaling alleviates vascular dysfunction in diabetic retinopathy
    Authors: JH Wu, YN Li, AQ Chen, CD Hong, CL Zhang, HL Wang, YF Zhou, PC Li, Y Wang, L Mao, YP Xia, QW He, HJ Jin, ZY Yue, B Hu
    EMBO Mol Med, 2020-01-13;12(2):e10154.
    Species: Mouse
    Sample Types: Protein, Whole Cells
    Applications: Functional Assay, Western Blot
  13. ADAM10 is Expressed by Ameloblasts, Cleaves the RELT TNF Receptor Extracellular Domain and Facilitates Enamel Development
    Authors: A Ikeda, S Shahid, BR Blumberg, M Suzuki, JD Bartlett
    Sci Rep, 2019-10-01;9(1):14086.
    Species: Mouse
    Sample Types: Protein
    Applications: Western Blot
  14. Phenotypic heterogeneity and evolution of melanoma cells associated with targeted therapy resistance
    Authors: Y Su, M Bintz, Y Yang, L Robert, AHC Ng, V Liu, A Ribas, JR Heath, W Wei
    PLoS Comput. Biol., 2019-06-05;15(6):e1007034.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  15. Organoid single cell profiling identifies a transcriptional signature of glomerular disease
    Authors: JL Harder, R Menon, EA Otto, J Zhou, S Eddy, NL Wys, C O'Connor, J Luo, V Nair, C Cebrian, JR Spence, M Bitzer, OG Troyanskay, JB Hodgin, RC Wiggins, BS Freedman, M Kretzler
    JCI Insight, 2019-01-10;4(1):.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  16. MicroRNA-149-5p regulates blood-brain barrier permeability after transient middle cerebral artery occlusion in rats by targeting S1PR2 of pericytes
    Authors: Y Wan, HJ Jin, YY Zhu, Z Fang, L Mao, Q He, YP Xia, M Li, Y Li, X Chen, B Hu
    FASEB J., 2018-01-18;0(0):fj201701121R.
    Species: Rat
    Sample Types: Cell Lysates, Whole Cells
    Applications: ICC, Western Blot
  17. BDNF/TrkB axis activation promotes epithelial-mesenchymal transition in idiopathic pulmonary fibrosis
    Authors: E Cherubini, S Mariotta, D Scozzi, R Mancini, G Osman, M D'Ascanio, P Bruno, G Cardillo, A Ricci
    J Transl Med, 2017-09-22;15(1):196.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  18. MMP20 modulates cadherin expression in ameloblasts as enamel develops.
    Authors: Guan X, Bartlett J
    2013-09-25;92(0):.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  19. In vivo biomarker expression patterns are preserved in 3D cultures of Prostate Cancer.
    Authors: Windus L, Kiss D, Glover T, Avery V
    Exp Cell Res, 2012-07-27;318(19):2507-19.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: ICC, Western Blot
  20. Rab coupling protein mediated endosomal recycling of N-cadherin influences cell motility
    Authors: Andrew J. Lindsay, Mary W. McCaffrey
    Oncotarget
  21. VPS34-dependent control of apical membrane function of proximal tubule cells and nutrient recovery by the kidney
    Authors: Markus M. Rinschen, Jennifer L. Harder, Madalina E. Carter-Timofte, Luis Zanon Zanon Rodriguez, Carmen Mirabelli, Fatih Demir et al.
    Science Signaling
  22. Distinct roles of VE ‐cadherin for development and maintenance of specific lymph vessel beds
    Authors: René Hägerling, Esther Hoppe, Cathrin Dierkes, Martin Stehling, Taija Makinen, Stefan Butz et al.
    The EMBO Journal
  23. Microenvironmental protection of CML stem and progenitor cells from tyrosine kinase inhibitors through N-cadherin and Wnt-beta-catenin signaling.
    Authors: Zhang B, Li M, McDonald T, Holyoake T, Moon R, Campana D, Shultz L, Bhatia R
    Blood, 2013-01-08;121(10):1824-38.

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