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7785
HTScan® VEGF Receptor 1 Kinase Assay Kit
Cellular Assay Kits
Assay Kit

HTScan® VEGF Receptor 1 Kinase Assay Kit #7785

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Figure 3. Dose dependence curve of VEGFR-1 kinase activity: DELFIA® data generated using Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 to detect phosphorylation of substrate peptide (#1310) by VEGFR-1 kinase. In a 50 µl reaction, increasing amounts of VEGFR-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 VEGFR-1 kinase activity: DELFIA® data generated using Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 to detect phosphorylation of VEGFR-1 substrate peptide (#1310) by VEGFR-1 kinase. In a 50 µl reaction, 50 ng VEGFR-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 VEGFR-1 kinase activity: DELFIA® data generated using Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 to detect phosphorylation of substrate peptide (#1310) by VEGFR-1 kinase. In a 50 µl reaction, 50 ng of VEGFR-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 VEGFR-1 kinase activity: DELFIA® data generated using Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 to detect phosphorylation of VEGFR-1 substrate peptide (#1310) by VEGFR-1 kinase. In a 50 µl reaction, 50 ng VEGFR-1 and 1.5 µM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)
Figure 1. VEGFR-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: PolyEY5 µg/50 µl, recombinant VEGFR-1: 100 ng/50 µl.
Inquiry Info.# 7785

Product Description

The kit provides a means of performing kinase activity assays with recombinant human VEGFR-1 kinase. It includes active VEGFR-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-Gastrin Precursor (Tlyr87): 2,853 Daltons. GST-VEGFR-1 Kinase: 89 kDa.

Background

The vascular endothelial growth factor (VEGF) receptor (VEGFR-1, Flt-1) is a 180 kDa receptor tyrosine kinase belonging to the VEGFR (Flt) family (1-3). The receptor is comprised of seven extracellular Ig-like domains, a single transmembrane region and cytoplasmic tail containing the active kinase domain (1,2). VEGFR-1 plays an important role in endothelial cell function and normal vascular development, as well as in hematopoietic function (2,3). VEGF-A is a high affinity ligand of VEGFR-1. VEGFR-1 also binds VEGF-B and PLGF (2). Ligand binding results in very little VEGFR-1 kinase activation, and VEGFR-1/VEGF-A binding negatively regulates VEGF function by diverting the growth factor from other functional VEGF receptors (3).

Pathways

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For Research Use Only. Not for Use in Diagnostic Procedures.
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