Catalog # EP2751

EphA4 (Tyr-779)[conserved site], phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
WB	1:1000
IHC	1:100

Size 100 μl

Species Reactivity Hu, Rt, Ms, Ck

MW 120 kDa

The Eph family of Receptor tyrosine kinases and their Ephrin ligands are important for cell positioning and morphogenesis during development. Eph receptors are classified into 10 EphA and 6 EphB receptors, which preferentially bind to the type A and type B ephrins, respectively. The EphA4 receptor can inhibit axon outgrowth and has roles in regulating axon projections during neural development. EphA4 signaling pathways require its kinase activity and involve binding and activation of Rho-GTPase guanine nucleotide-exchange factors (GEFs). EphA4 activation leads autophosphorylation of Tyr-596 and Tyr-602, and the conserved sites in EphA2 are required for binding to the GEFs, Vav2 and Vav3, and ephrin-induced cell migration. The Tyr-779 site in the kinase domain is also phosphorylated in vivo and may regulate kinase activity. Activated EphA4 leads to Src kinase phosphorylation of the GEF, ephexin-1, and this activates RhoA. Thus, EphA4 signaling involves complex tyrosine phosphorylation in its cytoplasmic region along with interaction with several GEFs.

References

Fang, W.B. et al. (2008) J. Biol. Chem. 283(23):16017.
Lackmann, M. & Boyd, A.W. (2008). Sci. Signal. 1(15):re2.
Binns, K.L. et al. (2000) Mol. Cell. Biol. 20(13):4791.

Western blot analysis of human umbilical vein endothelial cells untreated (lanes 1, 3, 5, & 7) or treated with pervanadate (1 mM) for 30 min. (lanes 2, 4, 6, & 8). The blot was probed with anti-EphA4 (N-terminal region) (lanes 1 & 2), anti-EphA4 (Tyr-779) (lanes 3 & 4), anti-EphA4 (Tyr-602) (lanes 5 & 6), or anti-EphA4 (C-terminal region) (lanes 7 & 8).

EphA4 (Tyr-779) synthetic peptide corresponds to amino acids surrounding phosphorylated tyrosine 779 in human EphA4. This sequence has significant homology to the conserved site in rat and mouse EphA4, and is conserved well in most other EphA family members, but has less conservation in EphB family members.

*For more information, see UniProt Accession P54764

Rabbit polyclonal, affinity-purified antibody is supplied in 100μl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

This antibody was cross-adsorbed to unrelated phosphotyrosine peptides before affinity purification using EphA4 (Tyr-779) peptide. The purified antibody detects a 120 kDa* band corresponding to EphA4 in western blots of HUVEC cells treated with pervanadate.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

Product References:

Hameetman, L. et al. (2015) Oncotarget. 6(31):31868. (WB/IHC: PC3 cells, Prostate carcinoma)

This kit contains: