Mouse EphB3 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF432

R&D Systems, part of Bio-Techne
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AF432
AF432-SP
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Conjugate
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Biotin
BAF432
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Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Mouse

Applications

Validated:

CyTOF-ready, Flow Cytometry, Immunocytochemistry, Western Blot

Cited:

Flow Cytometry, Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

View all EphB3 Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse EphB3
Leu30-Thr537
Accession # P54754

Specificity

Detects mouse EphB3 in direct ELISAs and Western blots. In direct ELISAs, approximately 5% cross-reactivity with recombinant human EphB3 is observed and less than 1% cross-reactivity with recombinant rat EphB1, recombinant mouse (rm) EphA4, rmEphB2, rmEphB4, and rmEphB6 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Mouse EphB3 Antibody

Detection of Mouse EphB3 antibody by Western Blot.

Detection of Mouse EphB3 by Western Blot.

Western blot shows lysates of mouse brain tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse EphB3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF432) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for EphB3 at approximately 130 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
EphB3 antibody in COLO 205 Human Cell Line by Immunocytochemistry (ICC).

EphB3 in COLO 205 Human Cell Line.

EphB3 was detected in immersion fixed COLO 205 human colorectal adeno-carcinoma cell line using Goat Anti-Mouse EphB3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF432) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; Catalog # NL001) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Applications for Mouse EphB3 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: COLO 205 human colorectal adenocarcinoma cell line

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed COLO 205 human colorectal adenocarcinoma cell line

Western Blot

1 µg/mL
Sample: Mouse brain tissue

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
Size / Price
Qty

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.

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 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 525 amino acid (aa) extracellular domain (ECD) of mature mouse 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, mouse EphB3 shares 96% and 99% aa sequence identity with human and rat EphB3, respectively. 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. Ruiz, J.C. et al. (1994) Mech. Dev. 48:153.
  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. Liu, X. et al. (2006) J. Neurosci. 26:3087.
  14. Batlle, E. et al. (2005) Nature 435:1126.
  15. Holmberg, J. et al. (2006) Cell 125:1151.

Long Name

Eph Receptor B3

Alternate Names

Cek10, Hek2, Mdk5, Sek4, Tyro6

Entrez Gene IDs

2049 (Human); 13845 (Mouse)

Gene Symbol

EPHB3

UniProt

P54754

Additional EphB3 Products

Product Documents for Mouse EphB3 Antibody

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Product Specific Notices for Mouse EphB3 Antibody

For research use only

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