N-Cadherin Antibody (13A9)

Immunocytochemistry: N-Cadherin Antibody (13A9) [NBP1-48309] - Immunostaining of original tumor, low passage, and high passage KCI-MENG1 cells. The original patient-derived tumor (top row) showed moderate and patchy immunoreactivity for epithelial membrane antigen (EMA); strong and diffuse immunostaining for progesterone receptor (PR); and a Ki-67 proliferative index of 2-3%. There was also strong immunostaining for N-cadherin and vimentin. KCI-MENG1-LP cells (middle row) and KCI-MENG1-HP cells (bottom row) maintained expression of EMA, N-cadherin, and vimentin but had significantly reduced PR expression compared to the original tumor. Whereas Ki-67 labeling was found in only a small number of cells in the original tumor and low passage cells, it was positive in virtually all P84 cells. Scale bar 50 um. Image collected and cropped by CiteAb from the following publication (//www.translational-medicine.com/content/13/1/227), licensed under a CC-BY licence.

Western Blot: N-Cadherin Antibody (13A9) [NBP1-48309] - Western blots showing a reduction in epithelial mesenchymal transition (EMT) markers upon PDK1 knockdown in Colo205 and HT29 cells using E cadherin (NBP2-19051), N cadherin (NBP1-48309) and B-actin antibody (NB600-501). The corresponding secondary antibodies used were either goat anti-rabbit IgG-HRP (NB7160) or goat anti-mouse IgG-HRP (NB7539). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33738242) licensed under a CC-BY licence.

Western Blot: N-Cadherin Antibody (13A9) [NBP1-48309] - Analysis of N-Cadherin expression in HeLa whole cell lysate.

Immunocytochemistry: N-Cadherin Antibody (13A9) [NBP1-48309] - Human meningioma mouse xenograft model KCI-MENG1-LPSX generated with the spontaneously immortal cell line KCI-MENG1-LP. Tumors from immunocompromised SCID mice were dissected and the derivative cell line KCI-MENG1-LPSX CL was generated. The EMA, PR, and N-cadherin IHC of the mouse tumor highly resembled the original patient-derived tumor. The vimentin- and Ki-67-stained cells in the mouse tumor tissue were markedly more abundant and more intensely stained than in the original tumor. KCI-MENG1-LPSX CL cells displayed the same patterns of immunostaining as the high passage parent cell line KCI-MENG1-HP, including the loss of PR staining. Scale bar 50 um. Image collected and cropped by CiteAb from the following publication (//www.translational-medicine.com/content/13/1/227), licensed under a CC-BY licence.

Flow (Intracellular): N-Cadherin Antibody (13A9) [NBP1-48309] - An intracellular stain was performed on HeLa with NBP1-48309 and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody.

Simple Western: N-Cadherin Antibody (13A9) [NBP1-48309] - Simple Western lane view shows a specific band for N Cadherin in 1.0 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunocytochemistry/Immunofluorescence: N-Cadherin Antibody (13A9) [NBP1-48309] - Adult mouse neural stem cells stained for N-Cadherin. Antibody at 1:100. ICC/IF image submitted by a verified customer review.

Immunohistochemistry-Paraffin: N-Cadherin Antibody (13A9) [NBP1-48309] - Analysis of a FFPE tissue section of human brain using 1:200 dilution of N-Cadherin antibody. The staining was developed using HRP labeled anti-mouse secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin.

Flow Cytometry: N-Cadherin Antibody (13A9) [NBP1-48309] - An intracellular stain was performed on U-251 MG cells with N-Cadherin Antibody (13A9) NBP1-48309AF594 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 594.

Immunocytochemistry/Immunofluorescence: N-Cadherin Antibody (13A9) [NBP1-48309] - HEK 293 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton X-100. The cells were incubated with anti- at 5 ug/mL overnight at 4C and detected with an anti-mouse DyLight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry: N-Cadherin Antibody (13A9) [NBP1-48309] - IHC analysis of N Cadherin in mouse liver using DAB with hematoxylin counterstain.

Flow Cytometry: N-Cadherin Antibody (13A9) [NBP1-48309] - An intracellular stain was performed on HeLa cells with NBP1-48309AF488 and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, Flow, Flow-IC, ICC/IF, IHC, IHC-P, IP, ICC/IF
• Clone: 13A9
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

N-Cadherin Antibody (13A9) Summary

• Immunogen: This N-Cadherin Antibody (13A9) was developed against the cytoplasmic domain of human N Cadherin [Swiss-Prot# P19022].
• Localization: Cell membrane; Single-pass type I membrane protein.
• Marker: Mesenchymal Cells Marker
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: CDH2
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Flow (Intracellular)
  • Flow Cytometry
  • Immunocytochemistry
  • Immunocytochemistry/Immunofluorescence 1:100
  • Immunohistochemistry 1:50-1:200
  • Immunohistochemistry-Paraffin 1:50-1:100
  • Immunoprecipitation 1:10-1:500
  • Simple Western 1:50
  • Western Blot 0.5 ug/ml
• Application Notes: In Western Blot a band is observed at approx. 140 kDa.
  
  In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
  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.
• Theoretical MW: 140 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.
• Control:

  HeLa Whole Cell Lysate

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Tris-Glycine, 0.15 M NaCl
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein G purified

Alternate Names for N-Cadherin Antibody (13A9)

• 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

Background

N-Cadherin, also referred to as Neural Cadherin (NCAD) or Cadherin-2 (CHD2), is a 130 kDa protein that is a member of the calcium-dependent adhesion molecule family of classical (type I) cadherins (1-4). Under the CDH2 gene, human N-cadherin is synthesized as a 906 amino acid protein with a theoretical molecular weight of 99.8 kDa (5). The N-cadherin protein structure is similar to other classical type I cadherins including epithelial (E-) cadherin and placental (P-) cadherin (1,2). N-cadherin consists of a 25 amino acid (aa) N-terminal signal peptide and 134 aa pro-peptide, a 565 aa extracellular domain (ECD) with five cadherin repeats, a 21 aa transmembrane segment, and a 161 aa cytoplasmic domain (1-3,5). The ECD of N-cadherin monomers is responsible for homotypic binding through either cis or trans adhesion (2,3).

N-cadherin is expressed on multiple cell types but is most highly expressed by mesenchymal cells and neural tissue (2). Functionally, N-cadherin has a number of roles including maintaining structural integrity and adhesion, cell signaling, and formation of neuronal synapses and the vascular wall (2). The cytoplasmic tail interacts with beta-catenin which then binds with alpha-catenin, forming the cadherin-catenin adhesion complex, an important component of adhesions junctions (1-3). Given its role in adhesion, N-cadherin serves as an indicator of epithelial-to-mesenchymal transition (EMT) (1-4). The loss of E-cadherin during EMT corresponds with an increase in N-cadherin expression (1-4). This "cadherin-switch" is associated with increased migratory and invasive behavior observed in tumor progress (1-4). Proteases including activity of a disintegrin and metalloprotease 10 (ADAM10), matrix metalloproteinases (MMPs), caspase 3, presenilin, and calpain can cleave N-cadherin as a mechanism for regulating Wnt/beta-catenin signaling and inducing oncogenic signals (3,4). In addition to its expression in solid tumors, N-cadherin has been indicated in hematological disorders such as leukemia and multiple myeloma (1). N-cadherin antagonists are currently being studied as potential therapeutics for a variety of cancer studies (1-2).

References

1. Mrozik, K. M., Blaschuk, O. W., Cheong, C. M., Zannettino, A., & Vandyke, K. (2018). N-cadherin in cancer metastasis, its emerging role in haematological malignancies and potential as a therapeutic target in cancer. BMC Cancer. https://doi.org/10.1186/s12885-018-4845-0

2. Loh, C. Y., Chai, J. Y., Tang, T. F., Wong, W. F., Sethi, G., Shanmugam, M. K., Chong, P. P., & Looi, C. Y. (2019). The E-Cadherin and N-Cadherin Switch in Epithelial-to-Mesenchymal Transition: Signaling, Therapeutic Implications, and Challenges. Cells. https://doi.org/10.3390/cells8101118

3. Derycke, L. D., & Bracke, M. E. (2004). N-cadherin in the spotlight of cell-cell adhesion, differentiation, embryogenesis, invasion and signalling. The International Journal of Developmental Biology. https://doi.org/10.1387/ijdb.041793ld

4. Yu, W., Yang, L., Li, T., & Zhang, Y. (2019). Cadherin Signaling in Cancer: Its Functions and Role as a Therapeutic Target. Frontiers in Oncology. https://doi.org/10.3389/fonc.2019.00989

5. Unitprot (P1903)

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 N-Cadherin Antibody (NBP1-48309)(38)

Publications using NBP1-48309

• Huang M, Wang C, Yao Y et al. Mebendazole-Induced Blood-Testis Barrier Injury in Mice Testes by Disrupting Microtubules in Addition to Triggering Programmed Cell Death International journal of molecular sciences Apr 11 2022 [PMID: 35457043]
• Mohammed S, Torres Luquis O, Walls E, Lloyd F. Lymph-Circulating Tumor Cells show distinct properties to Blood-Circulating Tumor Cells and constitute extraordinary efficient metastatic precursors bioRxiv Oct 15 2018 (ICC/IF)
• Karim S, Burzangi AS, Ahmad A et al. PI3K-AKT Pathway Modulation by Thymoquinone Limits Tumor Growth and Glycolytic Metabolism in Colorectal Cancer International journal of molecular sciences Feb 19 2022 [PMID: 35216429] (WB, Human)
• Matta R, Yousafzai MS, Murrell M, Gonzalez AL Endothelial cell secreted metalloproteinase-2 enhances neural stem cell N-cadherin expression, clustering, and migration FASEB journal : official publication of the Federation of American Societies for Experimental Biology Feb 1 2021 [PMID: 33417253]
• SAenz-de-Santa-MarIa I, Celada L, San JosE MartInez A et al. Blockage of Squamous Cancer Cell Collective Invasion by FAK Inhibition Is Released by CAFs and MMP-2 Cancers Dec 10 2020 [PMID: 33321813] (WB, Human)
• Beauchamp P, Jackson C B et al. 3D Co-culture of hiPSC-Derived Cardiomyocytes With Cardiac Fibroblasts Improves Tissue-Like Features of Cardiac Spheroids. Front Mol Biosci Mar 3 2020 [PMID: 32118040] (IHC, Mouse)
• Mohammed S, Torres Luquis O, Walls E, Lloyd F. Lymph-Circulating Tumor Cells show distinct properties to Blood-Circulating Tumor Cells and constitute extraordinary efficient metastatic precursors Acta Biomaterialia Oct 1 2018 [PMID: 31026363] (ICC/IF)
• Zhao, K;Li, X;Chen, X;Zhu, Q;Yin, F;Ruan, Q;Xia, J;Niu, Z; Inhibition of miR-140-3p or miR-155-5p by antagomir treatment sensitize chordoma cells to chemotherapy drug treatment by increasing PTEN expression Eur. J. Pharmacol. Apr 10 2019 [PMID: 30980798] (WB, Human)
• Hlavac N, VandeVord PJ. Astrocyte Mechano-Activation by High-Rate Overpressure Involves Alterations in Structural and Junctional Proteins Front Neurol Feb 22 2019 [PMID: 30853931] (WB, Rat)
• Ren W, Hou J, Yang C et al. Extracellular vesicles secreted by hypoxia pre-challenged mesenchymal stem cells promote non-small cell lung cancer cell growth and mobility as well as macrophage M2 polarization via miR-21-5p delivery J. Exp. Clin. Cancer Res. Feb 8 2019 [PMID: 30736829] (WB, Human)
• Chiba Y, He B, Yoshizaki K et al. The transcription factor AmeloD stimulates epithelial cell motility essential for tooth morphology J. Biol. Chem. Nov 30 2018 [PMID: 30504223] (IHC, Mouse)
• Park S, Kim H, Kim K et al. Sonic hedgehog signalling regulates the self-renewal and proliferation of skin-derived precursor cells in mice Cell Prolif. Aug 27 2018 [PMID: 30151845] (ICC/IF, Mouse)
• Chen CY, Melo E, Jakob P et al. N-Terminomics identifies HtrA1 cleavage of thrombospondin-1 with generation of a proangiogenic fragment in the polarized retinal pigment epithelial cell model of age-related macular degeneration. Matrix Biol. Mar 20 2018 [PMID: 29572155] (IHC)
• Melo E, Oertle P, Trepp C et al. HtrA1 Mediated Intracellular Effects on Tubulin Using a Polarized RPE Disease Model. EBioMedicine 2017 Dec 13 [PMID: 29269042] (IHC, Human)
• Becker A B, Qian J et al. Heart and Nervous System Pathology in Compound Heterozygous Friedreich Ataxia. J Neuropathol Exp Neurol Jan 8 2017 [PMID: 28789479] (IHC, Human)
• Farjood Farhad, Vargis Elizabeth Physical disruption of cell-cell contact induces VEGF expression in RPE cells Physical disruption of cell-cell contact induces VEGF expression in RPE cells. Mol Vis. 2017 Jul 17 [PMID: 28761317] (ICC, Human)
  
  Details:
  Citation using the DyLight 405 version of this antibody.
• Koeppen AH, Becker AB, Feustel PJ et al. The significance of intercalated discs in the pathogenesis of Friedreich cardiomyopathy. J Neurol Sci 2016 Aug 15 [PMID: 27423584]
• Michelhaugh SK, Guastella AR, Varadarajan K et al. Development of patient-derived xenograft models from a spontaneously immortal low-grade meningioma cell line, KCI-MENG1 J Transl Med. 2015 Jul 16 [PMID: 26174772] (ICC/IF, Human)
  
  Details:
  N-Cadherin antibody used for Immunocytochemistry on KCI-MENG1 cells or KCI-MENG1-LPSX cells, which are dissociated cells from second generation xenograft mouse tumor) at a dilution of 1:100.
• Chow AK, Ng L, Lam CS et al. The Enhanced Metastatic Potential of Hepatocellular Carcinoma (HCC) Cells with Sorafenib Resistance. PLoS One. 2013 Nov 11 [PMID: 24244338] (WB, Human)
• Maeda M, Johnson E, Mandal SH, Lawson KR, Keim SA, Svoboda RA, Caplan S, Wahl JK 3rd, Wheelock MJ, Johnson KR. Expression of inappropriate cadherins by epithelial tumor cells promotes endocytosis and degradation of E-cadherin via competition for p120(ctn). Oncogene25(33):4595-604. 2006 Aug 3. [PMID: 16786001] (WB, Human)
• Jaggi M, Nazemi T, Abrahams NA, Baker JJ, Galich A, Smith LM, Balaji KC. N-cadherin switching occurs in high Gleason grade prostate cancer. Prostate66(2):193-9. 2006 Feb 1. [PMID: 16173043] (IHC-P, Human)
• Wahl JK 3rd, Kim YJ, Cullen JM, Johnson KR, Wheelock MJ. N-cadherin-catenin complexes form prior to cleavage of the proregion and transport to the plasma membrane. J Biol Chem278(19):17269-76. 2003 May 9. [PMID: 12604612] (WB, IP, ICC/IF, Human)
• Van Aken EH, Papeleu P, De Potter P, Bruyneel E, Philippe J, Seregard S, Kvanta A, De Laey JJ, Mareel MM. Structure and function of the N-cadherin/catenin complex in retinoblastoma. Invest Ophthalmol Vis Sci43(3):595-602. 2002 Mar. [PMID: 11867572] (WB, IP, Flow, ICC/IF, Human)
• Reyes-Mugica M, Meyerhardt JA, Rzasa J, Rimm DL, Johnson KR, Wheelock MJ, Reale MA. Truncated DCC reduces N-cadherin/catenin expression and calcium-dependent cell adhesion in neuroblastoma cells. Lab Invest81(2):201-10. 2001 Feb. [PMID: 11232642] (WB, ICC/IF, Human)
• Kim JB, Islam S, Kim YJ, Prudoff RS, Sass KM, Wheelock MJ, Johnson KR. N-Cadherin extracellular repeat 4 mediates epithelial to mesenchymal transition and increased motility. J Cell Biol151(6):1193-206. 2000 Dec 11. [PMID: 11121435] (WB, ICC/IF, Human)
• Mialhe A, Levacher G, Champelovier P, Martel V, Serres M, Knudsen K, Seigneurin D. Expression of E-, P-, n-cadherins and catenins in human bladder carcinoma cell lines. J Urol164(3 Pt 1):826-35. 2000 Sep. [PMID: 10953163]
• Nieman MT, Prudoff RS, Johnson KR, Wheelock MJ. N-cadherin promotes motility in human breast cancer cells regardless of their E-cadherin expression. J Cell Biol;147(3):631-44. 1999 Nov 1. [PMID: 10545506] (WB, ICC/IF, Human)
• Makrigiannakis A, Coukos G, Christofidou-Solomidou M, Gour BJ, Radice GL, Blaschuk O, Coutifaris C. N-cadherin-mediated human granulosa cell adhesion prevents apoptosis: a role in follicular atresia and luteolysis? . Am J Pathol;154(5):1391-406. 1999 May. [PMID: 10329592] (WB, ICC/IF, Human)
• Benson DL, Tanaka H. N-cadherin redistribution during synaptogenesis in hippocampal neurons. J Neurosci18(17):6892-904. 1998 Sep 1. [PMID: 9712659] (WB, ICC/IF, Rat)
• Navarro P, Ruco L, Dejana E. Differential localization of VE- and N-cadherins in human endothelial cells: VE-cadherin competes with N-cadherin for junctional localization. J Cell Biol140(6):1475-84. 1998 Mar 23. [PMID: 9508779] (ICC/IF, Human)
• Peralta Soler A, Knudsen KA, Tecson-Miguel A, McBrearty FX, Han AC, Salazar H. Expression of E-cadherin and N-cadherin in surface epithelial-stromal tumors of the ovary distinguishes mucinous from serous and endometrioid tumors. Hum Pathol28(6):734-9. 1997 Jun. [PMID: 9191009]
• Han AC, Peralta-Soler A, Knudsen KA, Wheelock MJ, Johnson KR, Salazar H. Differential expression of N-cadherin in pleural mesotheliomas and E-cadherin in lung adenocarcinomas in formalin-fixed, paraffin-embedded tissues. Hum Pathol28(6):641-5. 1997 Jun. [PMID: 9190996] (Human)
• Hazan RB, Kang L, Whooley BP, Borgen PI. N-cadherin promotes adhesion between invasive breast cancer cells and the stroma. Cell Adhes Commun4(6):399-411. 1997 Mar. [PMID: 9177902]
• Islam S, Carey TE, Wolf GT, Wheelock MJ, Johnson KR. Expression of N-cadherin by human squamous carcinoma cells induces a scattered fibroblastic phenotype with disrupted cell-cell adhesion. J Cell Biol135(6 Pt 1):1643-54. 1996 Dec. [PMID: 8978829] (IP, WB, ICC/IF, Human)
• Peralta Soler A, Knudsen KA, JaurMC, Johnson KR, Wheelock MJ, Klein-Szanto AJ, Salazar H. The differential expression of N-cadherin and E-cadherin distinguishes pleural mesotheliomas from lung adenocarcinomas. Hum Pathol26(12):1363-9. 1995 Dec. [PMID: 8522310] (ICC/IF, Human)
• Knudsen KA, Soler AP, Johnson KR, Wheelock MJ. Interaction of alpha-actinin with the cadherin/catenin cell-cell adhesion complex via alpha-catenin. J Cell Biol130(1):67-77. 1995 Jul. [PMID: 7790378] (IP, WB, ICC/IF, Human)
• Sacco PA, McGranahan TM, Wheelock MJ, Johnson KR. Identification of plakoglobin domains required for association with N-cadherin and alpha-catenin. J Biol Chem270(34):20201-6. 1995 Aug 25. [PMID: 7650039] (WB, Human)

Reviews for N-Cadherin Antibody (NBP1-48309) (2)

Average Rating: 5

(Based on 2 reviews)

We have 2 reviews tested in 2 species: Human, Mouse.

reviewed by: Rita Matta Immunocytochemistry Mouse 07/23/2020

Summary

• Application: Immunocytochemistry
• Sample Tested: adult mouse neural stem cells
• Species: Mouse
• Lot: C-2

reviewed by: Chia-Wei Chang Western Blot Human 12/28/2011

Summary

• Application: Western Blot
• Sample Tested: Human
• Species: Human

Bioinformatics

• Gene Symbol: CDH2
• Entrez:
  • 12558(Mouse)
• Uniprot:
  • P19022()