Cell Signaling Technology

Product Pathways - Tyrosine Kinase/ Adaptors

Phospho-PDGF Receptor α (Tyr754) (23B2) Rabbit mAb #2992

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H M (R) Endogenous 198 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by Western blot.

Protocols

Specificity / Sensitivity

Phospho-PDGF Receptor α (Tyr754) (23B2) Rabbit mAb detects endogenous levels of PDGFRα only when phosphorylated at tyrosine 754. The antibody does not cross-react with activated PDGFRβ.

Source / Purification

Monoclonal antibodies are produced by immunizing rabbits with a synthetic phosphopeptide (KLH-coupled) corresponding to residues surrounding Tyr754 of human PDGFRα.

Western Blotting

Western Blotting

Phospho-PDGF Receptor α (Tyr754) (23B2) Rabbit mAb specifically binds to tyrosine phosphorylated PDGF receptor α, but not other phosphorylated tyrosine kinases. Western blot analysis of of extracts from cells expressing different activated tyrosine kinase proteins, using Phospho-PDGF Receptor α (Tyr754) (23B2) Rabbit mAb (upper) or Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 (lower).

Background

The proteins of the platelet derived growth factor (PDGF) family exist as several disulphide-bonded, dimeric isoforms (PDGF AA, PDGF AB, PDGF BB, PDGF CC and PDGF DD) that bind in a specific pattern to two closely related receptor tyrosine kinases, PDGF receptor α (PDGFRα) and PDGF receptor β (PDGFRβ). PDGFRα and PDGFRβ share 75% to 85% sequence homology between their two intracellular kinase domains while the kinase insert and carboxy-terminal tail regions display a lower level (27% to 28%) of homology (1). PDGF Receptor α homodimers bind all PDGF isoforms except those containing PDGF D. PDGF Receptor β homodimers bind PDGF BB and DD isoforms, as well as the PDGF AB heterodimer. The heteromeric PDGFα/β receptor binds PDGF B, C, and D homodimers as well as the PDGF AB heterodimer (2). PDGFRα and PDGFRβ can each form heterodimers with EGFR, which is also activated by PDGF (3). Various cells differ in the total number of receptors present and in the receptor subunit composition, which may account for responsive differences among cell types to PDGF binding (4). Ligand binding induces receptor dimerization and autophosphorylation, followed by binding and activation of cytoplasmic SH2 domain-containing signal transduction molecules such as Grb2, Src, GAP, PI3 kinase, PLCγ and Nck. A number of different signaling pathways are initiated by activated PDGF receptors and lead to control of cell growth, actin reorganization, migration and differentiation (5). Tyr751 in the kinase-insert region of PDGFRβ is the docking site for PI3 kinase (6). Phosphorylated pentapeptides derived from Tyr751 of PDGFRβ (pTyr751-Val-Pro-Met-Leu) inhibit the association of the carboxy-terminal SH2 domain of the p85 subunit of PI3 kinase with PDGFRβ (7). Tyr740 is also required for PDGFRβ mediated PI3 kinase activation (8).

Interestingly, PDGFR-alpha was found to be phosphorylated at an additional tyrosine residue, Tyr754, in a heterodimeric complex as compared to the alpha-alpha homodimer. Phosphorylation of this tyrosine residue permits the binding of a specific signal-transducing protein, and thereby initiates signaling pathway(s) from the beta-alpha heterodimer, which are dinstinct from those initiated via homodimeric receptor complexes (8).

  1. Deuel, T.F. et al. (1988) Biofactors 1, 213-217.
  2. Bergsten, E. et al. (2001) Nat. Cell Biol. 3, 512-516.
  3. Betsholtz, C. et al. (2001) Bioessays 23, 494-507.
  4. Coughlin, S.R. et al. (1988) Prog. Clin. Biol. Res. 266, 39-45.
  5. Ostman, A. and Heldin, C.H. (2001) Adv. Cancer Res. 80, 1-38.
  6. Panayotou, G. et al. (1992) EMBO J. 11, 4261-4272.
  7. Ramalingam, K. et al. (1995) Bioorg. Med. Chem. 3, 1263-1272.
  8. Kashishian, A. et al. (1992) EMBO J. 11, 1373-1382.
  9. Rupp, E. et al. (1994) Eur J Biochem 225, 29-41.
  10. Soroceanu, L. et al. (2008) Nature 455, 391-5.

Application References

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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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