EphB4 Antibody

EphB4 was detected in immersion fixed frozen sections of mouse embryo (15 d.p.c.) using Goat Anti-Mouse EphB4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF446) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

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

EphB4 Antibody Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse EphB4
  Leu16-Ala539
  Accession # P54761
• Specificity: Detects mouse EphB4 in direct ELISAs and Western blots. In direct ELISAs, approximately 5% cross-reactivity with recombinant mouse (rm) EphB6 and rmEphA3 is observed and less than 1% cross-reactivity with rmEphA2, rmEphA4, rmEphA6, rmEphA7, rmEphA8, rmEphB2, and rmEphB3 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: EPHB4
• Purity: Immunogen affinity purified
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 2.5 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
• Purity: Immunogen affinity purified
• 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 EphB4 Antibody

• EC 2.7.10
• EC 2.7.10.1
• EPH receptor B4
• EphB4
• ephrin type-B receptor 4
• hepatoma transmembrane kinase
• Htk
• HTKephrin receptor EphB4
• Mdk2
• Myk1
• soluble EPHB4 variant 1
• soluble EPHB4 variant 2
• soluble EPHB4 variant 3
• Tyro11
• Tyrosine-protein kinase receptor HTK
• Tyrosine-protein kinase TYRO11

Background

EphB4, also known as Htk, Myk1, Tyro11, and Mdk2 (1), is a member of the Eph receptor family which binds members of the ephrin ligand family. There are two classes of receptors, designated A and B. Both the A and B class receptors have an extracellular region consisting of a globular domain, a cysteine-rich domain, and two fibronectin type III domains. This is followed by the transmembrane region and cytoplasmic region. The cytoplasmic region contains a juxtamembrane motif with two tyrosine residues, which are the major autophosphorylation sites, a kinase domain, and a conserved sterile alpha motif (SAM) in the carboxy tail which contains one conserved tyrosine residue. Activation of kinase activity occurs after ligand recognition and binding. EphB4 has been shown to bind ephrin‑B2 and ephrin‑B1 (2, 3). The extracellular domains of human and mouse EphB4 share 88% amino acid identity. Only membrane-bound or Fc-clustered ligands are capable of activating the receptor in vitro. While soluble monomeric ligands bind the receptor, they do not induce receptor autophosphorylation and activation (2). In vivo, the ligands and receptors display reciprocal expression (3). It has been found that nearly all receptors and ligands are expressed in developing and adult neural tissue (3). The Eph/ephrin families also appear to play a role in angiogenesis (3).

1. Eph Nomenclature Committee [letter] (1997) Cell 90:403.
2. Flanagan, J.G. and P. Vanderhaeghen (1998) Annu. Rev. Neurosci. 21:309.
3. Pasquale, E.B. (1997) Curr. Opin. Cell Biol. 9:608.

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 EphB4 Antibody (AF446)(31)

Publications using AF446

• AE Lewis, A Kuwahara, J Franzosi, JO Bush Tracheal separation is driven by NKX2-1-mediated repression of Efnb2 and regulation of endodermal cell sorting Cell Reports, 2022;38(11):110510. 2022 [PMID: 35294885] (IF, Mouse)
• J Alam, K Ogawa Expression and localisation of ephrin-B1 and EphB4 in steroidogenic cells in the naturally cycling mouse ovary Reproductive biology, 2021;21(3):100511. 2021 [PMID: 33991765] (IHC, Mouse)
• C Donato, L Kunz, F Castro-Gin, A Paasinen-S, K Strittmatt, BM Szczerba, R Scherrer, N Di Maggio, W Heusermann, O Biehlmaier, C Beisel, M Vetter, C Rochlitz, WP Weber, A Banfi, T Schroeder, N Aceto Hypoxia Triggers the Intravasation of Clustered Circulating Tumor Cells Cell Rep, 2020;32(10):108105. 2020 [PMID: 32905777] (Western Blot, Human)
• G Luxán, J Stewen, N Díaz, K Kato, SK Maney, A Aravamudha, F Berkenfeld, N Nagelmann, HC Drexler, D Zeuschner, C Faber, H Schillers, S Hermann, J Wiseman, JM Vaquerizas, ME Pitulescu, RH Adams Endothelial EphB4 maintains vascular integrity and transport function in adult heart Elife, 2019;8(0):. 2019 [PMID: 31782728] (Western Blot, Human)
• S Payne, M Gunadasa-R, A Neal, AN Redpath, J Patel, KM Chouliaras, I Ratnayaka, N Smart, S De Val Regulatory pathways governing murine coronary vessel formation are dysregulated in the injured adult heart Nat Commun, 2019;10(1):3276. 2019 [PMID: 31332177] (IHC-Fr, Mouse)
• A Neal, S Nornes, S Payne, MD Wallace, M Fritzsche, P Louphrasit, RN Wilkinson, KM Chouliaras, K Liu, K Plant, R Sholapurka, I Ratnayaka, W Herzog, G Bond, T Chico, G Bou-Ghario, S De Val Venous identity requires BMP signalling through ALK3 Nat Commun, 2019;10(1):453. 2019 [PMID: 30692543] (IHC, Mouse)
• E Groppa, S Brkic, A Uccelli, G Wirth, P Korpisalo-, M Filippova, B Dasen, V Sacchi, MG Muraro, M Trani, S Reginato, R Gianni-Bar, S Ylä-Herttu, A Banfi EphrinB2/EphB4 signaling regulates non-sprouting angiogenesis by VEGF EMBO Rep., 2018;0(0):. 2018 [PMID: 29643120] (IHC-Fr, Mouse)
• H Toray, T Hasegawa, N Sakagami, E Tsuchiya, A Kudo, S Zhao, Y Moritani, M Abe, T Yoshida, T Yamamoto, T Yamamoto, K Oda, N Udagawa, PH Luiz de Fr, M Li Histochemical assessment for osteoblastic activity coupled with dysfunctional osteoclasts in c-src deficient mice Biomed. Res., 2017;38(2):123-134. 2017 [PMID: 28442663] (IHC, Mouse)
• EphrinB2 repression through ZEB2 mediates tumour invasion and anti-angiogenic resistance Nat Commun, 2016;7(0):12329. 2016 [PMID: 27470974] (Western Blot, Mouse)
• Downregulation of proinflammatory cytokines in HTLV-1-infected T cells by Resveratrol J Exp Clin Cancer Res, 2016;35(1):118. 2016 [PMID: 27448598] (IHC-P, Mouse)
• EPHB4 kinase-inactivating mutations cause autosomal dominant lymphatic-related hydrops fetalis J Clin Invest, 2016;0(0):. 2016 [PMID: 27400125] (IHC-Fr, Mouse)
• M Wang, MJ Collins, TR Foster, H Bai, T Hashimoto, JM Santana, C Shu, A Dardik Eph-B4 mediates vein graft adaptation by regulation of endothelial nitric oxide synthase J. Vasc. Surg., 2017;65(1):179-189. 2017 [PMID: 26817610] (Western Blot, Mouse)
• Nguyen T, Arthur A, Panagopoulos R, Paton S, Hayball J, Zannettino A, Purton L, Matsuo K, Gronthos S EphB4 Expressing Stromal Cells Exhibit an Enhanced Capacity for Hematopoietic Stem Cell Maintenance. Stem Cells, 2015;33(9):2838-49. 2015 [PMID: 26033476] (Western Blot, Mouse)
• Zhang , Gu, Brady , John, Liang , Wei-Chin, Wu , Yan, Henkemeyer , Mark, Yan , Minhong EphB4 forward signalling regulates lymphatic valve development. Nat Commun, 2015;6(0):6625. 2015 [PMID: 25865237] (Western Blot, Mouse)
• Valverde-Franco G, Hum D, Matsuo K, Lussier B, Pelletier J, Fahmi H, Kapoor M, Martel-Pelletier J The in vivo effect of prophylactic subchondral bone protection of osteoarthritic synovial membrane in bone-specific Ephb4-overexpressing mice. Am J Pathol, 2015;185(2):335-46. 2015 [PMID: 25453723] (IHC-P, Mouse)
• Hughes A, Spelke D, Xu Z, Kang C, Schaffer D, Herr A Single-cell western blotting. Nat Methods, 2014;11(7):749-55. 2014 [PMID: 24880876] (Western Blot, Rat)
• Lindskog , Henrik, Kim , Yung Hae, Jelin , Eric B, Kong , Yupeng, Guevara-Gallardo , Salvador, Kim , Tyson N, Wang , Rong A Molecular identification of venous progenitors in the dorsal aorta reveals an aortic origin for the cardinal vein in mammals. Development, 2014;141(5):1120-8. 2014 [PMID: 24550118] (IHC, Mouse)
• Lange A, Haitchi H, LeCras T, Sridharan A, Xu Y, Wert S, James J, Udell N, Thurner P, Whitsett J Sox17 is required for normal pulmonary vascular morphogenesis. Dev Biol, 2014;387(1):109-20. 2014 [PMID: 24418654]
• Kawano H, Katayama Y, Minagawa K, Shimoyama M, Henkemeyer M, Matsui T A novel feedback mechanism by Ephrin-B1/B2 in T-cell activation involves a concentration-dependent switch from costimulation to inhibition. Eur. J. Immunol., 2012;42(6):1562-72. 2012 [PMID: 22622783] (Immunoprecipitation, Mouse)
• Muto A, Yi T, Harrison KD, Davalos A, Fancher TT, Ziegler KR, Feigel A, Kondo Y, Nishibe T, Sessa WC, Dardik A Eph-B4 prevents venous adaptive remodeling in the adult arterial environment. J. Exp. Med., 2011;208(3):561-75. 2011 [PMID: 21339325] (IHC, Mouse)
• Foster KE, Gordon J, Cardenas K EphB-ephrin-B2 interactions are required for thymus migration during organogenesis. Proc. Natl. Acad. Sci. U.S.A., 2010;107(30):13414-9. 2010 [PMID: 20616004] (Flow Cytometry, Mouse)
• Wang Y, Nakayama M, Pitulescu ME, Schmidt TS, Bochenek ML, Sakakibara A, Adams S, Davy A, Deutsch U, Luthi U, Barberis A, Benjamin LE, Makinen T, Nobes CD, Adams RH Ephrin-B2 controls VEGF-induced angiogenesis and lymphangiogenesis. Nature, 2010;465(7297):483-6. 2010 [PMID: 20445537] (IHC-P, Mouse)
• Lin FJ, Chen X, Qin J, Hong YK, Tsai MJ, Tsai SY Direct transcriptional regulation of neuropilin-2 by COUP-TFII modulates multiple steps in murine lymphatic vessel development. J. Clin. Invest., 2010;120(5):1694-707. 2010 [PMID: 20364082] (IHC-P, Mouse)
• Xie J, Wu T, Xu K, Huang IK, Cleaver O, Huang CL Endothelial-specific expression of WNK1 kinase is essential for angiogenesis and heart development in mice. Am. J. Pathol., 2009;175(3):1315-27. 2009 [PMID: 19644017] (IHC-Fr, Mouse)
• Kim YH, Hu H, Guevara-Gallardo S, Lam MT, Fong SY, Wang RA Artery and vein size is balanced by Notch and ephrin B2/EphB4 during angiogenesis. Development, 2008;135(22):3755-64. 2008 [PMID: 18952909] (IHC-Fr, Mouse)
• Allan EH, Hausler KD, Wei T, Gooi JH, Quinn JM, Crimeen-Irwin B, Pompolo S, Sims NA, Gillespie MT, Onyia JE, Martin TJ EphrinB2 regulation by PTH and PTHrP revealed by molecular profiling in differentiating osteoblasts. J. Bone Miner. Res., 2008;23(8):1170-81. 2008 [PMID: 18627264] (Western Blot, Mouse)
• Zhang J, Dong H, Wang B, Zhu S, Croy BA Dynamic changes occur in patterns of endometrial EFNB2/EPHB4 expression during the period of spiral arterial modification in mice. Biol. Reprod., 2008;79(3):450-8. 2008 [PMID: 18463357] (IHC-P, Mouse)
• Yang NY, Pasquale EB, Owen LB, Ethell IM The EphB4 receptor-tyrosine kinase promotes the migration of melanoma cells through Rho-mediated actin cytoskeleton reorganization. J. Biol. Chem., 2006;281(43):32574-86. 2006 [PMID: 16950769] (Immunoprecipitation, Mouse)
• Ogawa K, Wada H, Okada N, Harada I, Nakajima T, Pasquale EB, Tsuyama S EphB2 and ephrin-B1 expressed in the adult kidney regulate the cytoarchitecture of medullary tubule cells through Rho family GTPases. J. Cell. Sci., 2006;119(0):559-70. 2006 [PMID: 16443753] (IHC, Mouse)
• Martiny-Baron G, Korff T, Schaffner F, Esser N, Eggstein S, Marme D, Augustin HG Inhibition of tumor growth and angiogenesis by soluble EphB4. Neoplasia, 2004;6(3):248-57. 2004 [PMID: 15153337] (Western Blot, Human)
• Fuller T, Korff T, Kilian A, Dandekar G, Augustin HG Forward EphB4 signaling in endothelial cells controls cellular repulsion and segregation from ephrinB2 positive cells. J. Cell. Sci., 2003;116(0):2461-70. 2003 [PMID: 12734395] (Western Blot, Human)

Bioinformatics

• Gene Symbol: EPHB4
• Uniprot:
  • P54761()