Phospho-Ephrin-B (Mouse/Rat Y328) Antibody

Catalog #: PPS048 Datasheet / COA / SDS

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PPS048

Detection of Phospho-Ephrin-B (Y328) by Western Blot.

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Phospho-Ephrin-B (Mouse/Rat Y328) Antibody Summary

Species Reactivity

Human, Mouse, Rat, Chicken, Xenopus

Specificity

This antibody is specific for the ~45 kDa Ephrin-B phosphorylated at Y328 in Western blots of rat testis.

Source

Polyclonal Rabbit IgG

Purification

Antigen Affinity-purified

Immunogen

Phosphopeptide corresponding to amino acid residues surrounding the phospho-Y317 of chicken Ephrin-B

Formulation

100 μL in 10 mM HEPES (pH 7.5), 150 mM NaCl, 100 μg/mL BSA and 50% glycerol.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1:1000 dilution

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 Phospho-Ephrin-B (Y328) by Western Blot. Western blot of rat testis lysate showing specific immunolabeling of the approximately 45 kDa Ephrin-B phosphorylated at Y328 (Control). The phosphospecificity of this labeling is shown in the second lane (lambda-phosphatase, lambda -Phosphatase). The blot is identical to the control except that it was incubated in lambda PPase (1200 units for 30 minutes) before being exposed to the anti-Ephrin-B (Y328). The immunolabeling of the Ephrin-B band is completely eliminated by treatment with lambda PPase.

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Preparation and Storage

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

For long-term storage, ≤ ‑20° C is recommended. Product is stable at ≤ ‑20° C for at least 1 year.

Background: Ephrin-B

Ephrin-B1 (from ephoros; Greek for controller) is a 45 kDa, type I transmembrane glycoprotein found on a number of highly divergent cell types. It is a ligand for the Eph family of receptor tyrosine kinases, principally EphB1, 2 and 3. It participates in angiogenesis, T cell development and activation, platelet adhesion, neural crest migration and axonal patterning. In rodent, Ephrin-B1 contains a 212 amino acid (aa) extracellular domain (ECD) and an 88 aa cytoplasmic region. The ECD exhibits a characteristic four Cystein pattern, while the cytoplasmic region displays a three aa C‑terminal PZD binding motif (Tyr-Lys-Val). Upon receptor ligation, Ephrin-B1 is known to be phosphorylated on tyrosine residues. Based on motifs that are seven amino acids in length, three tyrosines are conserved, Xenopus to chicken to human to rodent (mouse; rat). The first tyrosine is Xenopus Y298, chicken Y305, human Y317, and rodent Y316. This is contained in a common CPHYEKV motif. The second tyrosine is Xenopus Y310, chicken Y317, human Y329, and rodent Y328. This is contained in a common HPVYIVQ motif. The third tyrosine is Xenopus Y324, chicken Y331, human Y343, and rodent Y343 that is contained in a common ANIYYKV motif. Phosphorylation on rodent Y316 results in SH2-containing Grb4 interaction. Phosphorylation of rodent Y328 and Y343 may contribute to receptor oligomerization and/or interaction with trimeric G‑proteins. Based on Ephrin-B2 studies, rodent Y328 may also participate in SH2-domain binding.

References

Flanagan, J.C. and P. Vanderhaeghen (1998) Annu. Rev. Neurosci. 21:309.
Heroult, M. et al. (2006) Exp. Cell Res. 312:642.
Kalo, M.S. et al. (2001) J. Biol. Chem. 276:38940.
Bong, Y-S. et al. (2004) Biochem. J. 377:499.
Song, J. (2003) J. Biol. Chem. 278:24714.

Alternate Names

craniofrontonasal syndrome (craniofrontonasal dysplasia); EFL3; EFL-3; ELK ligand; Elk-L; EPH-related receptor tyrosine kinase ligand 2; EphrinB; Ephrin-B; ephrin-B1; EPLG2CFNS; LERK-2; LERK2CFND; ligand of eph-related kinase 2; MGC8782