HTScan® PKCι Kinase Assay Kit

• Supplier: Cell Signaling Technology
• Catalog: 7598
• Category: Assay Kit
• Description: The kit provides a means of performing kinase activity assays with recombinant human PKCiota kinase
• Gene: PRKCI

Product images

Figure 1. PKCiota 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), 0.5µg/50 µl and Recombinant PKCiota: 50 ng/50 µl.

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