Human EphB4 Antibody

(6 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human EphB4 in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross-reactivity with recombinant mouse (rm) EphB4 is observed and less than 1% cross-reactivity with recombinant human (rh) EphA4, rhEphA3, rhEphB2, and rhEphB3 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human EphB4
    Leu16-Ala539
    Accession # P54760
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    2 µg/mL
    See below
  • Simple Western
    50 µg/mL
    See below
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • Immunohistochemistry
    5-15 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.
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.
Data Examples
Detection of Human EphB4 by Western Blot. Western blot shows lysates of K562 human chronic myelogenous leukemia cell line, COLO 205 human colorectal adenocarcinoma cell line, ZR‑75 human breast cancer cell line, and HUVEC human umbilical vein endothelial cells. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human EphB4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3038) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for EphB4 at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of EphB4 in MCF‑7 Human Cell Line by Flow Cytometry. MCF‑7 human breast cancer cell line was stained with Goat Anti-Human EphB4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3038, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.
Immunohistochemistry
EphB4 in Human Kidney. EphB4 was detected in immersion fixed paraffin-embedded sections of human kidney using 15 µg/mL Goat Anti-Human EphB4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3038) overnight at 4 °C. Tissue was stained with 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 Paraffin-embedded Tissue Sections.
Detection of Human EphB4 by Simple WesternTM. Simple Western lane view shows lysates of HUVEC human umbilical vein endothelial cells, loaded at 0.2 mg/mL. A specific band was detected for EphB4 at approximately 127 kDa (as indicated) using 50 µg/mL of Goat Anti-Human EphB4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3038) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • 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: EphB4

EphB4, also known as Htk, Myk1, Tyro11, and Mdk2, is a member of the Eph receptor tyrosine kinase family and binds Ephrin-B2. The A and B class Eph proteins have a common structural organization (1-4). The human EphB4 cDNA encodes a 987 amino acid (aa) precursor that includes a 15 aa signal sequence, a 524 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 427 aa cytoplasmic domain (5). The ECD contains an N-terminal globular domain, a cysteine-rich domain, and two fibronectin type III domains. The cytoplasmic domain contains a juxtamembrane motif with two tyrosine residues which are the major autophosphorylation sites, a kinase domain, and a conserved sterile alpha motif (SAM) (5). Activation of kinase activity occurs after membrane-bound or clustered ligand recognition and binding. The ECD of human EphB4 shares 89% aa sequence identity with mouse EphB4 and 42-45% aa sequence identity with human EphB1, 2, and 3. EphB4 is expressed preferentially on venous endothelial cells (EC) and inhibits cell-cell adhesion, chemotaxis, and angiogenesis. Opposing effects are induced by signaling through Ephrin-B2 expressed on arterial EC: adhesion, endothelial cell migration, and vessel sprouting (6). EphB4 singaling contributes to new vascularization by guiding venous EC away from Ephrin-B2 expressing EC. Ephrin-B2 signaling induces arterial EC to migrate towards nascent EphB4 expressing vessels (6). The combination of forward signaling through EphB4 and reverse signaling through Ephrin-B2 promotes in vivo mammary tumor growth and
tumor-associated angiogenesis (7). EphB4 promotes the differentiation of megakaryocytic and erythroid progenitors but not granulocytic or monocytic progenitors (8, 9).

  • References:
    1. Poliakov, A. et al. (2004) Dev. Cell 7:465.
    2. Surawska, H. et al. (2004) Cytokine Growth Factor Rev. 15:419.
    3. Pasquale, E.B. (2005) Nat. Rev. Mol. Cell Biol. 6:462.
    4. Davy, A. and P. Soriano (2005) Dev. Dyn. 232:1.
    5. Bennett, B.D. et al. (1994) J. Biol. Chem. 269:14211.
    6. Fuller, T. et al. (2003) J. Cell Sci. 116:2461.
    7. Noren, N.K. et al. (2004) Proc. Natl. Acad. Sci. USA 101:5583.
    8. Wang, Z. et al. (2002) Blood 99:2740.
    9. Inada, T. et al. (1997) Blood 89:2757.
  • Long Name:
    Eph Receptor B4
  • Entrez Gene IDs:
    2050 (Human); 13846 (Mouse)
  • Alternate Names:
    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
Related Research Areas
Citations:

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.

6 Citations: Showing 1 - 6
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Species
Applications
Sample Type
  1. Downregulation of proinflammatory cytokines in HTLV-1-infected T cells by Resveratrol
    J Exp Clin Cancer Res, 2016;35(1):118.
    Species: Human
    Sample Type: Cell Lysates
    Application: IP
  2. EPHB4 kinase-inactivating mutations cause autosomal dominant lymphatic-related hydrops fetalis
    J Clin Invest, 2016;0(0):.
    Species: Human
    Sample Type: Cell Lysates
    Application: IP
  3. Instruction of circulating endothelial progenitors in vitro towards specialized blood-brain barrier and arterial phenotypes.
    Authors: Boyer-Di Ponio J, El-Ayoubi F, Glacial F, Ganeshamoorthy K, Driancourt C, Godet M, Perriere N, Guillevic O, Couraud P, Uzan G
    PLoS ONE, 2014;9(1):e84179.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC Fluorescence
  4. BCR-ABL-independent and RAS / MAPK pathway-dependent form of imatinib resistance in Ph-positive acute lymphoblastic leukemia cell line with activation of EphB4.
    Authors: Suzuki M, Abe A, Imagama S, Nomura Y, Tanizaki R, Minami Y, Hayakawa F, Ito Y, Katsumi A, Yamamoto K, Emi N, Kiyoi H, Naoe T
    Eur. J. Haematol., 2010;84(3):229-38.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  5. Soluble EphB4 inhibition of PDGF-induced RPE migration in vitro.
    Authors: He S, Kumar SR, Zhou P, Krasnoperov V, Ryan SJ, Gill PS, Hinton DR
    Invest. Ophthalmol. Vis. Sci., 2010;51(1):543-52.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Frozen
  6. Activation of the receptor EphB4 by its specific ligand ephrin B2 in human osteoarthritic subchondral bone osteoblasts.
    Authors: Kwan Tat S, Pelletier JP, Amiable N, Boileau C, Lajeunesse D, Duval N, Martel-Pelletier J
    Arthritis Rheum., 2008;58(12):3820-30.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Methylmethacrylate-embedde
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Isotype Controls
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Normal Goat IgG Control

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Secondary Antibodies
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Goat IgG (H+L) APC-conjugated Antibody

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Goat IgG Horseradish Peroxidase-conjugated Antibody

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Donkey Anti-Goat IgG Antibody

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Goat IgG (H+L) Fluorescein-conjugated Antibody

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Donkey Anti-Goat IgG NorthernLights™ NL637-conjugated Antibody

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Rabbit Anti-Goat IgG Biotinylated Antibody

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Chicken Anti-Goat IgG Biotinylated Antibody

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Donkey Anti-Goat IgG (H+L) PerCP-conjugated Antibody

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