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7737
HTScan® FGF Receptor 4 Kinase Assay Kit

HTScan® FGF Receptor 4 Kinase Assay Kit #7737

This product is discontinued

Kinase Assay - Radiometric Image 1

Figure 1. FGFR-4 kinase activity was measured in a radiometric assay using the following reaction conditions: 60 mM HEPES-NaOH, pH 7.5, 3 mM MgCl2, 3 mM MnCl2, 3 µM Na-orthovanadate, 1.2 mM DTT, ATP (variable), 2.5 µg/50 µl PEG20.000, Substrate: PolyEY, 2 µg/50 µl, recombinant FGFR-4: 50 ng/50 µl.

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Kinase Assay - DELFIA Image 2

Figure 3. Dose dependence curve of FGFR-4 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of substrate peptide (#1315) by FGFR-4 kinase. In a 50 µl reaction, increasing amounts of FGFR-4 and 1.5 µM substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

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Kinase Assay - DELFIA Image 3

Figure 5. Staurosporine inhibition of FGFR-4 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of FGFR-4 substrate peptide (#1315) by FGFR-4 kinase. In a 50 µl reaction, 50 ng FGFR-4, 1.5 µM substrate peptide, 20 µM ATP and increasing amounts of staurosporine were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

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Kinase Assay - DELFIA Image 4

Figure 4. Peptide concentration dependence of FGFR-4 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of substrate peptide (#1315) by FGFR-4 kinase. In a 50 µl reaction, 50 ng of FGFR-4 and increasing concentrations of substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

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Kinase Assay - DELFIA Image 5

Figure 2. Time course of FGFR-4 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of FGFR-4 substrate peptide (#1315) by FGFR-4 kinase. In a 50 µl reaction, 50 ng FGFR-4 and 1.5 µM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)

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Product Includes Quantity (with Count) Solution Color
DTT (1000X) 1 x 80 µl
ATP (10 mM) 9804 1 x 1 ml

The kit provides a means of performing kinase activity assays with recombinant human FGFR-4 kinase. It includes active FGFR-4 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-peptide: 2,166 Daltons. GST-FGFR-4 Kinase: 74 kDa.

Peptide Core Sequence:

DIY*AE

Fibroblast growth factors are a family of broad-spectrum growth factors influencing a plethora of cellular activities. The interaction of at least 23 ligands, 4 receptors and multiple coreceptors provides a dramatic complexity to a signaling system capable of effecting a multitude of responses (1,2). Basic fibroblast growth factor (bFGF or FGF2), initially identified as a mitogen with prominent angiogenic properties, is now recognized as a multifunctional growth factor (3). It is clear that bFGF produces its biological effects in target cells by signaling through cell-surface FGF receptors. bFGF binds to all four FGF receptors. Ligand binding induces receptor dimerization and autophosphorylation, allowing binding and activation of cytoplasmic downstream target proteins, including FRS-2, PLC and Crk (4,5). The FGF signaling pathway appears to play a significant role not only in normal cell growth regulation but also in tumor development and progression (6).

Acidic FGF (aFGF or FGF1) is another extensively investigated protein of the FGF family. aFGF shares 55% DNA sequence homology with bFGF. These two growth factors are ubiquitously expressed and exhibit a wide spectrum of similiar biological activities with quantitative differences likely due to variation in receptor affinity or binding (7).

  1. Powers, C.J. et al. (2000) Endocr Relat Cancer 7, 165-97.
  2. Bansal, R. (2002) Dev. Neurosci. 24, 35-46.
  3. Morrison, R.S. et al. (1994) J. Neurooncol. 18, 207-216.
  4. Kouhara, H. et al. (1997) Cell 89, 693-702.
  5. Mohammadi, M. et al. (1991) Mol Cell Biol 11, 5068-78.
  6. Bikfalvi, A. (1995) Eur. J. Cancer 31A, 1101-1104.
  7. Ledoux, D. et al. (1992) Prog. Growth Factor Res. 4, 107-120.
Entrez-Gene Id
2264
Swiss-Prot Acc.
P22455
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
DELFIA is a registered trademark of PerkinElmer, Inc.

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