Human EphB3 Antibody

Discontinued Product

AF5667 has been discontinued.
View all EphB3 products.
Detection of Human EphB3 by Western Blot.
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Human EphB3 Antibody Summary

Species Reactivity
Human
Specificity
Detects human EphB3 in direct ELISAs and Western blots. In direct ELISAs, approximately 3% cross-reactivity with recombinant mouse EphB3 is observed, and less than 1% cross-reactivity with recombinant rat EphB1, recombinant human (rh) EphB2 and rhEphB4 is observed.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human EphB3
Leu38-Ala550
Accession # P54753
Formulation
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below

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.

Scientific Data

Western Blot Detection of Human EphB3 antibody by Western Blot. View Larger

Detection of Human EphB3 by Western Blot. Western blot shows lysates of SH-SY5Y human neuroblastoma cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human EphB3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5667) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for EphB3 at approximately 110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Reconstitution Calculator

Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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: EphB3

EphB3, also known as Cek10, Tyro6, Sek4, Hek2, and Mdk5, is a 110-130 kDa member of the transmembrane Eph receptor tyrosine kinase family. The A and B classes of Eph proteins are distinguished by Ephrin ligand binding preference but have a common structural organization. Eph-Ephrin interactions are widely involved in the regulation of cell migration, tissue morphogenesis, and cancer progression (1). The 526 amino acid (aa) extracellular domain (ECD) of mature human EphB3 contains a ligand binding domain followed by a cysteine rich region and two fibronectin type III domains. The 418 aa cytoplasmic domain contains a tyrosine kinase domain, a sterile alpha motif (SAM), and a PDZ binding motif (2). Within the ECD, human EphB3 shares 96% aa sequence identity with mouse and rat EphB3. Binding of EphB3 to its ligands Ephrin-B1, B2, and B3 triggers forward signaling through EphB3 as well as reverse signaling through the Ephrin (1, 3). EphB3 also interacts in cis with the receptor tyrosine kinase Ryk (4). Activation of its kinase is required for some but not all of the effects of EphB3 on cellular adhesion, motility, and morphology (5). EphB3 is widely expressed during development and in the adult; it shows a complementary tissue distribution to the Ephrin-B ligands (6-9). EphB3 function is important in vascular, nervous system, thymocyte, and palate development (6, 7, 10-12). It directs embyronic neuronal axon pathfinding, and its upregulation on local macrophages following neuronal injury promotes the growth of regenerating axons (10, 13). EphB3 inhibits colorectal carcinogenesis and invasion by preventing the migration of tumor cells out of the intestinal crypt (9, 14). EphB3 function is supported by the cooperative action of EphB2 in several of these processes (6, 10-12, 15).

References
  1. Pasquale, E.B. (2008) Cell 133:38.
  2. Bohme, B. et al. (1993) Oncogene 8:2857.
  3. Pasquale, E.B (2004) Nat. Neurosci. 7:417.
  4. Trivier, E. and T.S. Ganesan (2002) J. Biol. Chem. 277:23037.
  5. Miao, H. et al. (2005) J. Biol. Chem. 280:923.
  6. Adams, R.H. et al. (1999) Genes Dev. 13:295.
  7. Krull, C.E. et al. (1997) Curr. Biol. 7:571.
  8. Willson, C.A. et al. (2006) J. Mol. Histol. 37:369.
  9. Cortina, C. et al. (2007) Nature Genet. 39:1376.
  10. Birgbauer, E. et al. (2000) Development 127:1231.
  11. Alfaro, D. et al. (2008) Immunology 125:131.
  12. Risley, M. et al. (2009) Mech. Dev. 126:230.
  13. 13. Liu, X. et al. (2006) J. Neurosci. 26:3087.
  14. 14. Batlle, E. et al. (2005) Nature 435:1126.
  15. 15. Holmberg, J. et al. (2006) Cell 125:1151.
Long Name
Eph Receptor B3
Entrez Gene IDs
2049 (Human); 13845 (Mouse)
Alternate Names
Cek10; EC 2.7.10; EC 2.7.10.1; EK2; EPH receptor B3; EphB3; EPH-like kinase 2; EPH-like tyrosine kinase-2; ephrin type-B receptor 3; ETK2Hek2; Hek2; human embryo kinase 2; Mdk5; Sek4; Tyro6; Tyrosine-protein kinase TYRO6

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Human EphB3 Antibody
By Anonymous on 09/23/2019
Application: ELISA Sample Tested: Serum and Plasma Species: Human

We used this antibody for developing a sandwich ELISA in combination with mAb (cat. MAB5667) and protein (cat. 5667 B3). This combination works well detecting Ephb3 in human serum.