HTScan® PDK1 Kinase Assay Kit

Name	HTScan® PDK1 Kinase Assay Kit
Supplier	Cell Signaling Technology
Catalog	7577
Category	Assay Kit
Description	The kit provides a means of performing kinase activity assays with recombinant human PDK1 kinase
Gene	PDPK1
Supplier Page	Shop

Product images

Figure 1. PDK1 kinase activity was measured in a radiometric assay using the following reaction conditions: 5 mM MOPS, pH 7.2, 2.5 mM β-glycerophosphate, 1 mM EGTA, 0.4 mM EDTA, 5 mM MgCl2, 0.05 mM DTT, 50 ng/μl BSA, 50 μM ATP, Substrate: PDKtide 400 ng/μL, and recombinant PDK1: variable.

Figure 3. Dose dependence curve of PDK1 kinase activity: DELFIA® data generated using Phospho-PKA C (Thr197) Antibody #4781 to detect phosphorylation of substrate peptide (#1024) by PDK1 kinase. In a 50 µl reaction, increasing amounts of PDK1 and 3.0 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 PDK1 kinase activity: DELFIA® data generated using Phospho-PKA C (Thr197) Antibody #4781 to detect phosphorylation of PDK1 substrate peptide (#1024) by PDK1 kinase. In a 50 µl reaction, 50 ng PDK1, 3.0 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 PDK1 kinase activity: DELFIA® data generated using Phospho-PKA C (Thr197) Antibody #4781 to detect phosphorylation of substrate peptide (#1024) by PDK1 kinase. In a 50 µl reaction, 50 ng of PDK1 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 PDK1 kinase activity: DELFIA® data generated using Phospho-PKA C (Thr197) Antibody #4781 to detect phosphorylation of PDK1 substrate peptide (#1024) by PDK1 kinase. In a 50 µl reaction, 50 ng PDK1 and 3.0 µM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)