Human EphB4 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human EphB4 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant human EphA1, A2, A5, A6, A10, B2, B3, B6, recombinant mouse EphA3, A4, or recombinant rat EphB1 is observed.
  • Source
    Monoclonal Rat IgG1 Clone # 395810
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human EphB4
    Ala16-Ala539
    Accession # AAH52804
  • 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
    1 µg/mL
    Recombinant Human EphB4 (Catalog # 3038-B4) under non-reducing conditions only
  • Flow Cytometry
    2.5 µg/106 cells
    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 EphB4 in MCF‑7 Human Cell Line by Flow Cytometry. MCF‑7 human breast cancer cell line was stained with Rat Anti-Human EphB4 Monoclonal Antibody (Catalog # MAB3038, filled histogram) or isotype control antibody (Catalog # MAB005, open histogram), followed by Allophycocyanin-conjugated Anti-Rat IgG F(ab')2 Secondary Antibody (Catalog # F0113).
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 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 precursor that includes a 15 amino acid (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. 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
Isotype Controls
Description Application Cat# Citations Images  

Rat IgG1 Isotype Control

Ctrl MAB005 37  
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Normal Rat IgG Control

Ctrl 6-001-A 9
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Normal Rat IgG Control (Azide Free)

Ctrl 6-001-F 1
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Rat IgG1 Isotype Control

Ctrl MAB005R
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Secondary Antibodies
Description Application Cat# Citations Images  

Rat IgG HRP-conjugated Antibody

WB, Simple Western HAF005 7  
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Goat Anti-Rat F(ab)2 IgG (H+L)
APC-conjugated Antibody

Flow F0113 2  
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Goat Anti-Rat IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL013 2
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Rat F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0105B 2  
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Goat Anti-Rat IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL015 1
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Goat Anti-Rat IgG Biotinylated Antibody

WB BAF005 2
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Rat IgG VisUCyte HRP Polymer Antibody

IHC VC005  
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Goat Anti-Rat IgG Antibody

WB AF005 1
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Rat F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0104B  
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Goat Anti-Rat IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL014
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Goat Anti-Rat F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0115 1
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