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7919
HTScan® Mouse PDGF Receptor α Kinase Assay Kit
Cellular Assay Kits
Assay Kit

HTScan® Mouse PDGF Receptor α Kinase Assay Kit #7919

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HTScan® Mouse PDGF Receptor α Kinase Assay Kit: Image 1
Western blot analysis of extracts from Jurkat cells treated with 1 mM pervanadate for 30 minutes prior to lysis, using P-Tyr-100 Phospho-Tyrosine Mouse mAb. Proteins were separated by 2-D electrophoresis prior to blotting.
HTScan® Mouse PDGF Receptor α Kinase Assay Kit: Image 2
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® Mouse PDGF Receptor α Kinase Assay Kit: Image 3
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® Mouse PDGF Receptor α Kinase Assay Kit: Image 4
Immunohistochemical analysis of paraffin-embedded NCI-H1650 xenograft untreated (left) or lambda-phosphatase-treated (right), using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® Mouse PDGF Receptor α Kinase Assay Kit: Image 5
Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin lymphoma, untreated (left) or lambda phosphatase treated (right), using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® Mouse PDGF Receptor α Kinase Assay Kit: Image 6
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® Mouse PDGF Receptor α Kinase Assay Kit: Image 7
Immunohistochemical analysis using Phospho-Tyrosine Mouse mAb (P-Tyr-100) on SignalSlide™ Phospho-EGF Receptor IHC Controls #8102 (paraffin-embedded KYSE450 cell pellets, untreated (left) or EGF-treated (right)).
HTScan® Mouse PDGF Receptor α Kinase Assay Kit: Image 8
Immunohistochemical analysis of paraffin-embedded human soft tissue squamous cell carcinoma using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® Mouse PDGF Receptor α Kinase Assay Kit: Image 9
Immunohistochemical analysis of paraffin-embedded normal human kidney using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® Mouse PDGF Receptor α Kinase Assay Kit: Image 10
Immunohistochemical analysis of paraffin-embedded normal human lung using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® Mouse PDGF Receptor α Kinase Assay Kit: Image 11
Immunohistochemical analysis of paraffin-embedded NCI-H1650 xenograft using Phospho-Tyrosine Mouse mAb (P-Tyr-100).
HTScan® Mouse PDGF Receptor α Kinase Assay Kit: Image 12
Confocal immunofluorescent analysis of paraffin-embedded human lung adenocarcinoma using Phospho-Tyrosine Mouse mAb (P-Tyr-100) (red). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
HTScan® Mouse PDGF Receptor α Kinase Assay Kit: Image 13
Immunofluorescent analysis of Swiss NIH/3T3 cells, serum-starved and stimulated with lysophosphatidic acid (LPA) (10 µM for 10 minutes) and fixed with PFA, using Phospho-Tyrosine Mouse mAb (P-Tyr-100) (red) and phalloidin for F-actin (green) LPA causes heavy tyrosine phosphorylation of proteins in focal adhesions, present at the tips of actin stress fibers. (Provided by Dr. Harry Mellor, University of Bristol.)
HTScan® Mouse PDGF Receptor α Kinase Assay Kit: Image 14
Confocal immunofluorescence analysis of HeLa cells either stimulated with 20% serum (left) or untreated (right), using Phospho-Tyrosine Mouse mAb (P-Tyr-100) (red). Actin filaments have been labeled with fluorescein phalloidin. Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
HTScan® Mouse PDGF Receptor α Kinase Assay Kit: Image 15
Flow cytometric analysis of K562 cells, untreated (green) or Gleevec®- treated (blue), using Phospho-Tyrosine Mouse mAb (P-Tyr-100) compared to a nonspecific negative control antibody (red).
Inquiry Info.# 7919

Product Description

The kit provides a means of performing kinase activity assays with recombinant mouse PDGFRalpha kinase. It includes active PDGFRalpha kinase (supplied as a GST fusion protein), a biotinylated peptide substrate and a phospho-tyrosine antibody for detection of the phosphorylated form of the substrate peptide.
Molecular Formula Peptide substrate, Biotin-FLT3 (Tyr589): 1,945 Daltons. GST-mouse PDGFRalpha Kinase domain: 87 kDa.

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.

Pathways & Proteins

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