Revision 1
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

190

Source/Isotype:

Rabbit IgG

UniProt ID:

#P16234

Entrez-Gene Id:

5156

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Phospho-PDGF Receptor α (Tyr762) (D9B1N) Rabbit mAb recognizes endogenous levels of PDGF Receptor α protein only when phosphorylated at Tyr762. The antibody might cross react slightly with other overexpressed phosphorylated receptor tyrosine kinase such as EGFR.

Species Reactivity:

Human

Species predicted to react based on 100% sequence homology

Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr762 of human PDGF Receptor α protein.

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).
Phosphorylation of PDGFRα at Tyr762 was identified at Cell Signaling Technology using PTMScan®, our LC-MS/MS platform for phosphorylation site discovery (6). Tyr762 is located in the activation loop of the PDGFRα kinase domain. Phosphorylation of PDGFRα at this site was also reported by several other labs to be a docking site for CrkII and CrkL upon induction by growth factor treatment (7,8).

  1. Deuel, T.F. et al. (1988) Biofactors 1, 213-7.
  2. Bergsten, E. et al. (2001) Nat Cell Biol 3, 512-6.
  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. Rikova, K. et al. (2007) Cell 131, 1190-203.
  7. Matsumoto, T. et al. (2000) Biochem Biophys Res Commun 270, 28-33.
  8. Yokote, K. et al. (1998) Oncogene 16, 1229-39.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PTMScan is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#24188

Phospho-PDGF Receptor α (Tyr762) (D9B1N) Rabbit mAb

Western Blotting Image 1: Phospho-PDGF Receptor α (Tyr762) (D9B1N) Rabbit mAb Expand Image
Western blot analysis of extracts from NCI-H1703 cells serum-starved overnight and then either untreated (-) or treated with Human Platelet-Derived Growth Factor AA (hPDGF-AA) #8913 (100 ng/ml, 10 min;+), using Phospho-PDGF Receptor α (Tyr762) Rabbit mAb (upper) or PDGF Receptor α (D13C6) XP® Rabbit mAb #5241 (lower).
Immunoprecipitation Image 1: Phospho-PDGF Receptor α (Tyr762) (D9B1N) Rabbit mAb Expand Image
Immunoprecipitation of Phospho-PDGF Receptor α (Tyr762) protein from Human Platelet-Derived Growth Factor AA (hPDGF-AA) #8913 treated (100 ng/ml, 5 min) NCI-H1703 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-PDGF Receptor α (Tyr762) (D9B1N) Rabbit mAb. Western blot analysis was performed using Phospho-PDGF Receptor α (Tyr762) (D9B1N) Rabbit mAb (multiple partially degraded phospho-PDGF Receptor α bands shown at lower molecular size).