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Anti-EphB4 (extracellular) Antibody

Ephrin type-B receptor 4, Eph receptor B4, Hepatoma transmembrane kinase, HTK
Cat #: AER-024
Alternative Name Ephrin type-B receptor 4, Eph receptor B4, Hepatoma transmembrane kinase, HTK
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide (C)HEKGAEGPSSVRFLK, corresponding to amino acid residues 472-486 of mouse EphB4 (Accession P54761). Extracellular, N-terminus.
Accession (Uniprot) Number P54761
Gene ID 13846
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Rat, human – identical.
RRID AB_2756584.
Purity Affinity purified on immobilized antigen.
Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Isotype Rabbit IgG.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution 0.8 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
Standard quality control of each lot Western blot analysis.
Applications: if, ifc, ih, wb
May also work in: ic*, ip*, lci*
Western blot
  • Western blot analysis of mouse (lanes 1 and 3) and rat (lanes 2 and 4) brain lysates:
    Western blot analysis of mouse (lanes 1 and 3) and rat (lanes 2 and 4) brain lysates:
    1,2. Anti-EphB4 (extracellular) Antibody (#AER-024), (1:200).
    3,4. Anti-EphB4 (extracellular) Antibody, preincubated with EphB4 (extracellular) Blocking Peptide (#BLP-ER024).
  • Western blot analysis of human THP-1 monocytic leukemia cell line lysate (lanes 1 and 4), human Jurkat T-cell leukemia cell line lysate (lanes 2 and 5) and human MCF-7 breast adenocarcinoma cell line lysate (lanes 3 and 6):
    Western blot analysis of human THP-1 monocytic leukemia cell line lysate (lanes 1 and 4), human Jurkat T-cell leukemia cell line lysate (lanes 2 and 5) and human MCF-7 breast adenocarcinoma cell line lysate (lanes 3 and 6):
    1-3. Anti-EphB4 (extracellular) Antibody (#AER-024), (1:200).
    4-6. Anti-EphB4 (extracellular) Antibody, preincubated with EphB4 (extracellular) Blocking Peptide (#BLP-ER024).
Immunohistochemistry
  • Expression of EphB4 in rat hippocampus
    Expression of EphB4 in rat hippocampus
    Immunohistochemical staining of perfusion-fixed frozen rat brain sections with Anti-EphB4 (extracellular) Antibody (#AER-024), (1:100), followed by goat anti-rabbit-AlexaFluor-488. EphB4 immunoreactivity (green) in the hippocampal dentate gyrus is detected in the granule layer (G) and hilar interneurons (arrows). Cell nuclei are stained with DAPI (blue).
Indirect flow cytometry
  • Cell surface detection of EphB4 in live intact human THP-1 monocytic leukemia cells:
    Cell surface detection of EphB4 in live intact human THP-1 monocytic leukemia cells:
    ___ Cells.
    ___
    Cells + goat-anti-rabbit-FITC.
    ___ Cells + Anti-EphB4 (extracellular) Antibody (#AER-024), (2.5 µg) + goat-anti-rabbit-FITC.
  • Cell surface detection of EphB4 in live intact human Jurkat T-cell leukemia cells:
    Cell surface detection of EphB4 in live intact human Jurkat T-cell leukemia cells:
    ___ Cells.
    ___
    Cells + goat-anti-rabbit-FITC.
    ___ Cells + Anti-EphB4 (extracellular) Antibody (#AER-024), (5 µg) + goat-anti-rabbit-FITC.
References
  1. Liu, Y. et al. (2018) PLoS One 13, e0198291.
  2. Cheng, C. et al. (2017) Exp. Eye Res. 162, 9.
  3. Merchant, A.A. et al. (2017) Blood Adv. 1, 1635.
Scientific background

EphB4 (Ephrin type-B receptor 4) is a member of the Eph family of receptor tyrosine kinases. Signaling pathways of these receptors play important roles in regulating numerous physiological processes during development including cell morphology, proliferation, migration and axonal guidance, dendritic spine formation and neural plasticity. Ephrins transduce bidirectional signals based on their distribution on the communicating cell surfaces. Eph receptors mediate forward signaling in one cell while ephrin ligands transmit reverse signaling in an adjacent cell1,2.

Eph receptors consist of two large subfamilies: EphAs and EphBs that bind to either glycophosphatidylinositol (GPI)-anchored ephrin-A ligands or transmembrane ephrin-B ligands respectively. Each eph receptor may interact with multiple ligands, except for EPHB4, which only binds ephrin-B22.

EPHB4 is expressed primarily during embryonic development in neural tissue and immature vasculature. Studies show that EphB4 is implicated in the growth of several epithelial tumors and is a promising target in cancer therapy. EphB4 signaling was found to promote tumor growth, invasiveness, chemoresistance and tumor angiogenesis3.

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat
Last update: 23/08/2020

Anti-EphB4 (extracellular) Antibody (#AER-024) is a highly specific antibody directed against an epitope of the mouse protein. The antibody can be used in western blot, immunohistochemistry, and live cell flow cytometry applications. It has been designed to recognize EphB4 from rat, mouse, and human samples.

For research purposes only, not for human use
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