Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-PDGF Receptor β (Tyr1009) (42F9) Rabbit mAb #3124

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H M Endogenous 190 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-PDGF Receptor β (Tyr1009) (42F9) Rabbit mAb detects endogenous levels of PDGF receptor β only when phosphorylated at Tyr1009. The antibody may slightly cross-react with other activated PDGF receptor family members and other activated protein tyrosine kinases.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1009 of human PDGF receptor β.

Western Blotting

Western Blotting

Western blot analysis of cell extracts from NIH/3T3 cells unstimulated or stimulated with PDGF-BB (100 ng/ml for 5 min), using Phospho-PDGF Receptor-β (Tyr1009) (42F9) Rabbit mAb (upper) or PDGF receptor-β (2B3) Mouse mAb #3175 (lower).

Background

Platelet derived growth factor (PDGF) family proteins 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). PDGFRα homodimers bind all PDGF isoforms except those containing PDGF D. PDGFRβ homodimers bind PDGF BB and DD isoforms, as well as the PDGF AB heterodimer. The heteromeric PDGF receptor α/β 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).

Activation of the PDGFRβ leads to autophosphorylation on a number of tyrosine residues, including Tyr1009. Mutation analysis has shown that PDGF-stimulated PLCγ signaling is dependent on autophosphorylation of the PDGFRβ at Tyr1009 and Tyr1021 (9). Phosphorylated Tyr1009 also serves as a binding site for SHP-2, a SH2 domain-containing tyrosine phosphatase that is tyrosine-phosphorylated by PDGFRβ (10).

  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. Rönnstrand, L. et al. (1992) EMBO J. 11, 3911-3919.
  10. Rönnstrand, L. et al. (1999) Oncogene 18, 3696-3702.

Application References

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Companion Products

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

For Research Use Only. Not For Use In Diagnostic Procedures.

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