Figure 1. PDGFRalpha kinase activity was measured in a radiometric assay using the following reaction conditions: 60 mM HEPES-NaOH, pH 7.5, 5 mM MgCl2, 5 mM MnCl2, 3 µM Na-orthovanadate, 1.2 mM DTT, 100 µM ATP, 100 µM FLT3 (Tyr589) Biotinylated Peptide (#1305) and variable amount of Recombinant PDGFRalpha. Reaction mixture incubated at room temperature for 10 minutes.
Figure 3. Dose dependence curve of PDGFRalpha kinase activity: DELFIA® data generated using Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 to detect phosphorylation of substrate peptide (#1305) by PDGFRalpha kinase. In a 50 µl reaction, increasing amounts of PDGFRalpha 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 PDGFRalpha kinase activity: DELFIA® data generated using Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 to detect phosphorylation of PDGFRalpha substrate peptide (#1305) by PDGFRalpha kinase. In a 50 µl reaction, 100 ng PDGFRalpha, 1.5 µM substrate peptide, 5 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 PDGFRalpha kinase activity: DELFIA® data generated using Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 to detect phosphorylation of substrate peptide (#1305) by PDGFRalpha kinase. In a 50 µl reaction, 100 ng of PDGFRalpha 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 PDGFRalpha kinase activity: DELFIA® data generated using Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 to detect phosphorylation of PDGFRalpha substrate peptide (#1305) by PDGFRalpha kinase. In a 50 µl reaction, 100 ng PDGFRalpha and 1.5 µM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)