HTScan® PKCζ Kinase Assay Kit

• Supplier: Cell Signaling Technology
• Catalog: 7607
• Category: Assay Kit
• Description: The kit provides a means of performing kinase activity assays with recombinant human PKCzeta kinase
• Gene: PRKCZ

Product images

Figure 1. PKCzeta kinase activity was measured in a radiometric assay using the following reaction conditions: 60 mM HEPES-NaOH, pH 7.5, 5 mM MgCl2, 1.32 mM CaCl2, 1 mM EDTA, 1.25 mM EGTA, 0.25 µg/50 µl phosphatidylserine, 50 ng/50 µl 1.2 Dioleyl-glycerol, 1.2 mM DTT, ATP: variable , 2.5 µg/50 µl PEG20.000, Substrate: PKC(19-31) (RFARKGSLRQKNV), 1.0 µg/50 µl and Recombinant PKCzeta: 200 ng/50 µl.

Figure 3. Dose dependence curve of PKCzeta kinase activity: DELFIA® data generated using Phospho-PKA Substrate (RRXS/T) (100G7) Rabbit mAb #9624 to detect phosphorylation of substrate peptide (#1331) by PKCzeta kinase. In a 50 µl reaction, increasing amounts of PKCzeta 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 PKCzeta kinase activity: DELFIA® data generated using Phospho-PKA Substrate (RRXS/T) (100G7) Rabbit mAb #9624 to detect phosphorylation of PKCzeta substrate peptide (#1331) by PKCzeta kinase. In a 50 µl reaction, 50 ng PKCzeta, 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 PKCzeta kinase activity: DELFIA® data generated using Phospho-PKA Substrate (RRXS/T) (100G7) Rabbit mAb #9624 to detect phosphorylation of substrate peptide (#1331) by PKCzeta kinase. In a 50 µl reaction, 50 ng of PKCzeta 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 PKCzeta kinase activity: DELFIA® data generated using Phospho-PKA Substrate (RRXS/T) (100G7) Rabbit mAb #9624 to detect phosphorylation of PKCzeta substrate peptide (#1331) by PKCzeta kinase. In a 50 µl reaction, 50 ng PKCzeta and 1.5 uM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)