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Anti-Ephrin-B2 (extracellular) Antibody

EFNB2, EPH-Related Receptor Tyrosine Kinase Ligand 5, LERK5, HTK ligand, HTKL

Cat #: AER-042
Alternative Name EFNB2, EPH-Related Receptor Tyrosine Kinase Ligand 5, LERK5, HTK ligand, HTKL
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide (C)RRPELEAGTNGRS, corresponding to amino acid residues 181 - 193 of human Ephrin-B2 (Accession P52799). Extracellular, N-terminus.
Accession (Uniprot) Number P52799
Gene ID 1948
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Mouse, rat – identical
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).
Preadsorption Control 1 µg peptide per 1 µg antibody
Ephrin-B2 (extracellular) Blocking Peptide (#BLP-ER042)
Standard quality control of each lot Western blot analysis.
Applications: ih, wb
May also work in: ic*, ip*, lci*
Western blot
  • Western blot analysis of rat brain lysate (lanes 1 and 3) and mouse brain lysate (lanes 2 and 4):
    Western blot analysis of rat brain lysate (lanes 1 and 3) and mouse brain lysate (lanes 2 and 4):
    1-2. Anti-Ephrin-B2 (extracellular) Antibody (#AER-042), (1:400).
    3-4. Anti-Ephrin-B2 (extracellular) Antibody, preincubated with Ephrin-B2 (extracellular) Blocking Peptide (BLP-ER042).
  • Western blot analysis of human Colo-205 colon carcinoma cell line lysate (lanes 1 and 3) and human U-87 MG glioblastoma cell line lysate (lanes 2 and 4):
    Western blot analysis of human Colo-205 colon carcinoma cell line lysate (lanes 1 and 3) and human U-87 MG glioblastoma cell line lysate (lanes 2 and 4):
    1-2. Anti-Ephrin-B2 (extracellular) Antibody (#AER-042), (1:400).
    3-4. Anti-Ephrin-B2 (extracellular) Antibody, preincubated with Ephrin-B2 (extracellular) Blocking Peptide (BLP-ER042).
Immunohistochemistry
  • Expression of Ephrin-B2 in rat hippocampus.
    Expression of Ephrin-B2 in rat hippocampus.
    Immunohistochemical staining of perfusion-fixed frozen rat brain sections with Anti-Ephrin-B2 (extracellular) Antibody (#AER-042), (1:200), followed by goat anti-rabbit-AlexaFluor-488. A. Staining in the hippocampal dentate gyrus region, showed immunoreactivity (green) in astrocytes of the hilus (H, arrows) and in neurons of the granule layer (G). B. Pre-incubation of the antibody with Ephrin-B2 (extracellular) Blocking Peptide (BLP-ER042), suppressed staining. Cell nuclei are stained with DAPI (blue).
References
  1. Tanaka, M. et al. (2007) J. Cell Sci120, 2179.
  2. Kataoka, H. et al. (2002) J. Cancer Res. Clin. Oncol. 128, 343.
  3. Pasquale, E.B. (2005) Mol. Cell Biol. 6, 462.
  4. Wang, H.U. et al (1998) Cell 93:741-53.
  5. Chen, Y. et al. (2012) Cell. Signal. 24, 606.
  6. DeBuysscher, Blair L et al. (2021) Cells vol. 10,6.
  7. Liu, Wei et al. (2022) Frontiers in cell and dev. Biol. 10,788587.
Scientific background

Ephrin-B2 is a ligand for RTK Eph receptors, mediating cell adhesion and repulsion-segregation, thus influencing cell migration and tissue morphogenesis and angiogenesis. Ephrin-B2 is highly expressed in the nervous system during development and required for neuronal survival. Also expressed in the brain and lung endothelial cells (EC) for angiogenesis. It is overexpressed in various cancers.

The erythropoietin-producing hepatocellular (Eph) receptors represent the largest known family of receptor tyrosine kinases and are activated by interactions with cell-surface ligands, termed ephrins. Eph receptors have been classified into two subfamilies, EphA and EphB, according to their preference for either glycosylphosphatidylinositol (GPI)-anchored ephrin-A ligands (ephrin-A1 to -A5) or transmembrane ephrin-B ligands (ephrin-B1 to -B3). All ephrin ligands share a conserved core sequence of approximately 125 amino acids, including 4 invariant cysteine residues, probably corresponding to a receptor binding domain 1.

Ephrin-B2 is a cell surface transmembrane ligand 2. Ephrin-B2 binds Eph receptors residing on adjacent cells, leading to contact-dependent bidirectional signaling into neighboring cells. The signaling pathway downstream of the receptor is referred to as forward signaling while the signaling pathway downstream of the ephrin ligand is referred to as reverse signaling. Forward and reverse signaling through Eph receptors is a unique characteristic to this RTK since ephrins are physically linked to the plasma membrane 3. For example, in angiogenesis EphB4-mediated forward signaling controls cellular repulsion and segregation from ephrin-B2-expressing cells, resulting in distinction between EphB4-expressing veins and ephrin-B2-expressing arteries 4.

Eph receptors and ephrins are highly expressed in the brain and in the developing nervous system, where they are involved in fundamental developmental processes of the nervous system, including axon guidance, axon fasciculation, neural crest cell migration, acquisition of brain subregional identity, and neuronal cell survival. They are also implicated in Alzheimer’s disease, and other neurodegenerative disorders 5.

Nipah virus (NiV) infection has both respiratory and encephalitic components and several cell types are targets of NiV, dictated by the expression of the ephrin-B2/3 ligand on the cell’s outer membrane, which interact with the NiV surface proteins. ephrin-B2 and ephrin-B3 on cells, are the only identified receptors for NiV and Hendra virus (HeV) from the family Paramyxoviridae. Vascular endothelial cells (ECs) are the major targets of infection by NiV and HeV 6 .

In cancer cells, Eph receptors and ephrins are overexpressed. Eph receptor bidirectional signaling with both tumor-promoting and tumor-suppressing activities has been observed in different cancer types and their microenvironments. EphB2 is aberrantly expressed in many cancer types, such as colorectal cancer, gastric cancer and hepatocellular carcinoma, driving tumorigenesis and tumor progression 7.

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: 08/01/2023

Anti-Ephrin-B2 (extracellular) Antibody (#AER-042) is a highly specific antibody directed against an extracellular epitope of the human protein. The antibody can be used in western blot and immunohistochemistry. It has been designed to recognize Ephrin-B2 from mouse, rat and human samples.

For research purposes only, not for human use
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