539672 PKCδ, Human, Recombinant, S. frugiperda

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539672
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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.
      Quality LevelMQ100
      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.