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7420
HTScan® FGF Receptor 1 Kinase Assay Kit

HTScan® FGF Receptor 1 Kinase Assay Kit #7420

This product is discontinued

Kinase Assay - Radiometric Image 1

Figure 1. FGFR-1 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, 5 µg/50 µl, recombinant FGFR-1: 50 ng/50 µl.

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Kinase Assay - DELFIA Image 2

Figure 3. Dose dependence curve of FGFR-1 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of substrate peptide (#1315) by FGFR-1 kinase. In a 50 µl reaction, increasing amounts of FGFR-1 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-1 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of FGFR-1 substrate peptide (#1315) by FGFR-1 kinase. In a 50 µl reaction, 50 ng FGFR-1, 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-1 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of substrate peptide (#1315) by FGFR-1 kinase. In a 50 µl reaction, 50 ng of FGFR-1 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-1 kinase activity: DELFIA® data generated using Phospho-Tyrosine mAb (P-Tyr-100) #9411 to detect phosphorylation of FGFR-1 substrate peptide (#1315) by FGFR-1 kinase. In a 50 µl reaction, 50 ng FGFR-1 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-1 kinase. It includes active FGFR-1 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-Pyk2 (Tyr402): 2,166 Daltons. GST-FGFR-1 Kinase: 78 kDa.

Peptide Core Sequence:

DIY*AE

Fibroblast growth factors (FGFs) produce mitogenic and angiogenic effects in target cells by signaling through the cellular surface tyrosine kinase receptors-FGF receptors. There are four members of the FGF receptor family: FGFR-1 (flg), FGFR-2 (bek, KGFR), FGFR-3 and FGFR-4. Each receptor contains an extracellular ligand binding domain, a transmembrane region and a cytoplasmic kinase domain (1). Following ligand binding and dimerization, the receptors are phosphorylated and the receptor tyrosine kinase activated (2), resulting in further activation of various downstream signaling molecules and lysosomal degradation. FGFR has peptide substrate specificity similar to other receptor tyrosine kinase members, prefering acidic residues at the -1 to -4 positions and large hydrophobic amino acids at positions +1 and +3 (3).

  1. Powers, C.J. et al. (2000) Endocr Relat Cancer 7, 165-97.
  2. Mohammadi, M. et al. (1996) Mol Cell Biol 16, 977-89.
  3. Songyang, Z. and Cantley, L.C. (1995) TIBS 20, 470-475.
Entrez-Gene Id
2260
Swiss-Prot Acc.
P11362
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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