Human EphB2 Antibody

Name: Human EphB2 Antibody
Brand: R&D Systems
SKU: MAB5189

Catalog #: MAB5189 Datasheet / COA / SDS

Discontinued Product

MAB5189 has been discontinued.

View all EphB2 products.

Detection of Human EphB2 by Western Blot.

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All EphB2 Antibodies

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human EphB2 Antibody Summary

Species Reactivity

Human

Specificity

Detects human EphB2 in direct ELISAs. In Western blots, less than 5% cross-reactivity with recombinant human (rh) EphA1, 2, 5, 6, 10, rhEphB3, 4, 6, recombinant mouse (rm) EphA3, 4, 7, 8, rmEphB2, or recombinant rat EphB1 is observed.

Source

Monoclonal Mouse IgG_2A Clone # 669025

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Mouse myeloma cell line NS0-derived recombinant human EphB2
Val19-Leu543
Accession # P29323

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.

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

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Detection of Human EphB2 by Western Blot. Western blot shows lysates of HEK293 human embryonic kidney cell line transfected with human EphB2, HeLa human cervical epithelial carcinoma cell line, NCI-H460 human large cell lung carcinoma cell line, Saos-2 human osteosarcoma cell line, SK-Mel-28 human malignant melanoma cell line, and U2OS human osteosarcoma cell line. PVDF Membrane was probed with 1 µg/mL of Human EphB2 Monoclonal Antibody (Catalog # MAB5189) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for EphB2 at approximately 105 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it 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: EphB2

EphB2, also known as Cek5, Nuk, Erk, Qek5, Tyro5, Sek3, Hek5, and Drt, is a 125 kDa member of the transmembrane Eph receptor tyrosine kinase family that binds members of the Ephrin family on adjacent cells. The interaction triggers forward signaling in the receptor-expressing cells through the Eph receptor and reverse signaling in the ligand-expressing cells through Ephrin (1, 2). Human EphB2 cDNA encodes a 1055 amino acid (aa) precursor, which includes an 18 aa signal sequence, a 525 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 491 aa cytoplasmic domain. The ECD contains a cysteine-rich region followed by two fibronectin type III domains. The cytoplasmic domain contains the tyrosine kinase domain, a sterile alpha motif (SAM), and a PDZ binding motif (3). Human EphB2 shares 99% aa sequence identity with mouse and rat EphB2 within the ECD region. A short isoform that lacks 70 aa at the C-terminus has also been reported (4). Hippocampal neurons can release vesicles containing full length EphB2, and these are taken up by neighboring glial cells (5). EphB2 is expressed on both sides of the neuronal synapse. It controls axon guidance across the embryonic midline, promotes a neuronal fate from neural precursors, and regulates NMDA receptor activity (6 10). EphB2 interaction with Ephrin A5 promotes axonal growth cone collapse, while its interaction with Ephrin B ligands is required for inner ear, renal, urorectal, and vascular development (6, 11 15). Signaling in Ephrin-expressing cells through EphB2-Ephrin complex requires proteolytic cleavage of EphB2 that releases its extracellular domain (16). Following the shedding of the extracellular domain of EphB2, the cytoplasmic domain of EphB2 is released from the plasma membrane by the presenilin-dependent gamma -secretase activity to initiate a signaling cascade in the EphB2-expressing cells (16). Aberrant EphB2 expression and activity are implicated in the progression of several cancers (17).

References

1. Pasquale, E.B. (2008) Cell 133:38.
2. MerlosSuarez, A. and E. Batlle (2008) Curr. Opin. Cell Biol. 20:194.
3. Fox, G.M. et al. (1995) Oncogene 10:897.
4. Tang, X.X. et al. (1998) Oncogene 17:521.
5. Lauterbach, J. and R. Klein (2006) J. Neurosci. 26:11575.
6. Cowan C.A. et al. (2000) Neuron 26:417.
7. Bouvier, D. et al. (2008) J. Neurochem. 106:682.
8. Cramer, K.S. et al. (2006) Dev. Biol. 295:76.
9. Katakowski, M. et al. (2005) Neurosci. Lett. 385:204.
10. Henderson, J.T. et al. (2001) Neuron 32:1041.
11. Himanen, J.P.et al. (2004) Nat. Neurosci. 7:501.
12. Dravis, C. et al. (2007) Hear. Res. 223:93.
13. Dravis, C. et al. (2004) Dev. Biol. 271:272.
14. Ogawa, K. et al. (2006) J. Cell Sci. 119:559.
15. Salvucci, O. et al. (2006) Blood 108:2914.
16. Litterst, C. et al. (2007) J. Biol. Chem. 282:16155.
17. Castano, J. et al. (2008) Histol. Histopathol. 23:1011.

Long Name

Eph Receptor B2

Entrez Gene IDs

2048 (Human); 13844 (Mouse)

Alternate Names

CAPB; Cek5; Drt; DRTEphB2; EC 2.7.10; EC 2.7.10.1; EK5; elk-related tyrosine kinase; EPH receptor B2; eph tyrosine kinase 3; EphB2; EPH-like kinase 5; ephrin type-B receptor 2; EPHT3MGC87492; EPTH3; Erk; ERKHek5; Hek5; Nuk; PCBC; protein-tyrosine kinase HEK5; Qek2; Renal carcinoma antigen NY-REN-47; Sek3; Tyro5; Tyrosine-protein kinase receptor EPH-3; Tyrosine-protein kinase TYRO5