Revision 3

#9360Store at -20C

1 Kit

(4 x 20 microliters)

Cell Signaling Technology

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

Support: 877-678-TECH (8324)

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Phospho-PDGF Receptor α (Tyr754) (23B2) Rabbit mAb 2992 20 µl 190 kDa Rabbit 
Phospho-PDGF Receptor α (Tyr849)/PDGF Receptor β (Tyr857) (C43E9) Rabbit mAb 3170 20 µl 190 kDa Rabbit IgG
Phospho-PDGF Receptor α (Tyr1018) Antibody 4547 20 µl 190 kDa Rabbit 
PDGF Receptor α (D1E1E) XP® Rabbit mAb 3174 20 µl 190 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The PDGF Receptor α Antibody Sampler Kit provides an economical means of evaluating total PDGF receptor α protein (PDGFRα) levels as well as PDGFRα phosphorylated at specific sites. The kit contains enough primary and secondary antibody to perform two western blots with each antibody.

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.

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).

  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.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a 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 3
    #9360

    PDGF Receptor α Antibody Sampler Kit

    PDGF Receptor α Antibody Sampler Kit: Image 1 Expand Image
    Immunoprecipitation of PDGF Receptor α from NCI H1703 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is PDGF Receptor α (D1E1E) XP® Rabbit mAb.
    PDGF Receptor α Antibody Sampler Kit: Image 2 Expand Image
    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).
    PDGF Receptor α Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of NIH/3T3 cells, untreated or treated with PDGFbb using Phospho-PDGF Receptor α (Tyr849)/PDGF Receptor β (Tyr857) (C43E9) Rabbit mAb (upper) and PDGF Receptor β (28E1) Rabbit mAb #3169 (lower).
    PDGF Receptor α Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from NIH/3T3 and human skeletal muscle cells (SKMC), untreated or treated with PDGF-BB, using PDGF Receptor α (D1E1E) XP® Rabbit mAb.
    PDGF Receptor α Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of cell extracts from NIH/3T3 cells, untreated or treated with PDGF-AA, using Phospho-PDGF Receptor α (Tyr1018) Antibody (upper) or PDGF Receptor α Antibody #3164 (lower).
    PDGF Receptor α Antibody Sampler Kit: Image 6 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    PDGF Receptor α Antibody Sampler Kit: Image 7 Expand Image
    Immunohistochemical analysis of paraffin-embedded human glioblastoma using PDGF Receptor α (D1E1E) XP® Rabbit mAb.
    PDGF Receptor α Antibody Sampler Kit: Image 8 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon using PDGF Receptor α (D1E1E) XP® Rabbit mAb.
    PDGF Receptor α Antibody Sampler Kit: Image 9 Expand Image
    Immunohistochemical analysis of paraffin-embedded U-118 MG xenograft using PDGF Receptor α (D1E1E) XP® Rabbit mAb in the presence of control peptide (left) or antigen specific peptide (right).
    PDGF Receptor α Antibody Sampler Kit: Image 10 Expand Image
    Immunohistochemical analysis of paraffin-embedded HCC827 xenograft using PDGF Receptor α (D1E1E) XP® Rabbit mAb.
    PDGF Receptor α Antibody Sampler Kit: Image 11 Expand Image
    Confocal immunofluorescent analysis of A-204 (left) and U-87 MG cells (right) using PDGF Receptor α (D1E1E) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    PDGF Receptor α Antibody Sampler Kit: Image 12 Expand Image
    Flow cytometric analysis of fixed and permeabilized Jurkat cells (blue, negative) and IMR-32 cells (green, positive) using PDGF Receptor α (D1E1E) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.