539672 PKCδ, Human, Recombinant, S. frugiperda

539672
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      Overview

      Replacement Information

      Pricing & Availability

      Catalog Number AvailabilityPackaging Qty/Pack Price Quantity
      539672-5UG
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          Plastic ampoule 5 μg
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          Description
          OverviewFull-length, recombinant, human PKCδ expressed in S. frugiperda insect cells using a baculovirus expression system. Suitable for studies of PKCδ and its regulation. PKCδ is a Ca2+-independent and phospholipid-dependent enzyme.
          Catalogue Number539672
          Brand Family Calbiochem®
          SynonymsProtein Kinase Cδ
          References
          ReferencesHug, J. and Sarre, T.F. 1993. Biochem. J. 291, 329.
          Kazanietz, M.G., et al. 1993. Mol. Pharmacol. 44, 298.
          Koide, H., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 1149.
          Product Information
          Unit of DefinitionOne unit is defined as the amount of enzyme that will transfer 1 nmol of phosphate to PKC<sub>ε</sub> Peptide Substrate (<a href ="/Products/ProductDisplay.asp→catNO=539562">Cat. No. 539562</a>) per min at 30°C, pH 7.5.
          FormLiquid
          FormulationIn 250 mM NaCl, 20 mM HEPES, 5 mM DTT, 2 mM EDTA, 2 mM EGTA, 50% glycerol, 0.05% TRITON® X-100 Detergent, pH 7.5.
          Applications
          Biological Information
          Purity≥95% by SDS-PAGE
          Specific Activity≥800 units/mg protein
          Concentration Label Please refer to vial label for lot-specific concentration
          Physicochemical Information
          Dimensions
          Materials Information
          Toxicological Information
          Safety Information according to GHS
          Safety Information
          Product Usage Statements
          Storage and Shipping Information
          Ship Code Dry Ice Only
          Toxicity Standard Handling
          Storage ≤ -70°C
          Avoid freeze/thaw Avoid freeze/thaw
          Do not freeze Ok to freeze
          Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
          Packaging Information
          Transport Information
          Supplemental Information
          Specifications

          Documentation

          PKCδ, Human, Recombinant, S. frugiperda SDS

          Title

          Safety Data Sheet (SDS) 

          PKCδ, Human, Recombinant, S. frugiperda Certificates of Analysis

          TitleLot Number
          539672

          References

          Reference overview
          Hug, J. and Sarre, T.F. 1993. Biochem. J. 291, 329.
          Kazanietz, M.G., et al. 1993. Mol. Pharmacol. 44, 298.
          Koide, H., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 1149.

          Brochure

          Title
          PKC Pathway Poster PDF ( 676 KB )
          Data Sheet

          Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

          Revision08-September-2008 RFH
          SynonymsProtein Kinase Cδ
          DescriptionFull-length, recombinant, human PKCδ expressed in S. frugiperda insect cells using a baculovirus expression system. PKC δ is classified as a novel PKC isozyme because it does not exhibit calcium dependence and it is activated in vitro by phorbol esters. Suitable for studies of PKCδ and its regulation. PKCδ is a Ca2+-independent and phospholipid-dependent enzyme.
          FormLiquid
          FormulationIn 250 mM NaCl, 20 mM HEPES, 5 mM DTT, 2 mM EDTA, 2 mM EGTA, 50% glycerol, 0.05% TRITON® X-100 Detergent, pH 7.5.
          Concentration Label Please refer to vial label for lot-specific concentration
          Recommended reaction conditions
          Protocol for Recombinant PKC δ Note: this preparation is purified to near homogeneity, so it may behave differently than crude preparations. In particular, the concentration and quality of the phospholipids used does not seem to be important. Lipid Mixture: 1. Determine the total amount of lipid mixture needed. Each reaction requires 20 µg of phosphatidylserine (1.2 µl of 10 mg/ml phosphatidylserine in CHCl3) and 2 mg of diacylglycerol (0.6 µl of 2 mg/ml diacylglycerol in CHCl3). Preparing 10% more lipid mixture than required is recommended to account for losses while pipetting. 2. Using a Hamilton syringe that has been cleaned with methanol, transfer the required volume of each lipid stock solution to a 12 x 75 mm glass tube. 3. Evaporate off the chloroform. 4. Resuspend in 10 µl of 10 mM HEPES, pH 7.4, 0.3% TRITON® X-100 detergent. 5. Vortex for at least 2 min. 6. Place the lipid mixture in a 40°C water bath for 5 min prior to adding the reaction mixture. Reaction Mixture: 2.4 µl 500 mM HEPES, pH 7.4 6 µl 100 mM MgCl2 6 µl 1 mM EGTA 6 µl 1 mg/ml PKC ε peptide (Cat. No. 539562) 0.6 µl 10 mM ATP 6 µl lipid mixture 0.1 µl γ-ATP (5 mCi/ml) 32.9 µl distilled H2O Total volume should be 60 µl. Prepare a sufficient reaction mixture for all assays to be performed. Reaction: 1. Dispense 60 µl of reaction mixture into each tube and incubate at 30°C. 2. Dilute the PKC δ stock solution (500 ng/µl) to between 20-50 ng/µl in 10 mM Tris-HCl, pH 7.5, 5 mM DTT, and 0.01% TRITON®: X-100 detergent. 3. Add 2 µl of diluted enzyme to each tube. 4. Incubate for 10 min. 5. Stop the reaction by adding 6 µl of 5% phosphoric acid. 6. Incubate on ice for 5 min. 7. Spot 55 µl from each tube to phosphocellulose membranes and allow to dry. 8. Wash the membranes 3X with 5 ml of 0.5% phosphoric acid per filter. 9. Transfer membranes to a scintillation vial and count. Note: This protocol is meant to serve as a guideline and may need to be modified for specific applications.
          Purity≥95% by SDS-PAGE
          Specific activity≥800 units/mg protein
          Unit definitionOne unit is defined as the amount of enzyme that will transfer 1 nmol of phosphate to PKCε Peptide Substrate (Cat. No. 539562) per min at 30°C, pH 7.5.
          Storage ≤ -70°C
          Avoid freeze/thaw
          Do Not Freeze Ok to freeze
          Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
          Toxicity Standard Handling
          ReferencesHug, J. and Sarre, T.F. 1993. Biochem. J. 291, 329.
          Kazanietz, M.G., et al. 1993. Mol. Pharmacol. 44, 298.
          Koide, H., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 1149.