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7420
HTScan® FGF Receptor 1 Kinase Assay Kit
Cellular Assay Kits

HTScan® FGF Receptor 1 Kinase Assay Kit #7420

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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.)

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.)

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.)

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.)

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.

To Purchase # 7420

Product Description

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

Background

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) Trends Biochem Sci 20, 470-5.

Pathways & Proteins

Explore pathways + proteins related to this product.

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.