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539660 PKCε, Human, Recombinant, S. frugiperda

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

Synonyms: Protein Kinase Cε

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539660
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Overview

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539660-20UGCN
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      		 Plastic ampoule 20 μg
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      Description
      OverviewFull-length, recombinant, human, PKCε expressed in S. frugiperda insect cells using a baculovirus expression system. Suitable for studies of PKCε and its regulation. PKCε is a Ca2+-independent and phospholipid-dependent enzyme.
      Catalogue Number539660
      Brand Family Calbiochem®
      SynonymsProtein Kinase Cε
      References
      ReferencesNewton, A.C. 1995. J. Biol. Chem. 270, 28495.
      Dekker, L.V., and Parker, P.J. 1994. Trends Biochem. Sci. 19, 73.
      Nishizuka, Y. 1992. Science 258, 607.
      Bell, R.M., and Burns, D.J. 1991. J. Biol. Chem. 266, 4661.
      Product Information
      Unit of DefinitionOne unit is defined as the amount of enzyme that will transfer 1 nmol 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 7.4.
      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 7.4.
      Quality LevelMQ100
      Applications
      Biological Information
      Purity≥85% 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
      Global Trade Item Number
      Catalogue Number GTIN
      539660-20UGCN 04055977194197

      Documentation

      PKCε, Human, Recombinant, S. frugiperda SDS

      Title

      Safety Data Sheet (SDS) 

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

      TitleLot Number
      539660

      References

      Reference overview
      Newton, A.C. 1995. J. Biol. Chem. 270, 28495.
      Dekker, L.V., and Parker, P.J. 1994. Trends Biochem. Sci. 19, 73.
      Nishizuka, Y. 1992. Science 258, 607.
      Bell, R.M., and Burns, D.J. 1991. J. Biol. Chem. 266, 4661.
      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.

      Revision12-May-2008 JSW
      SynonymsProtein Kinase Cε
      DescriptionFull-length, recombinant, human, PKCε expressed in S. frugiperda insect cells using a baculovirus expression system. Suitable for studies of PKCε and its regulation. PKCε 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 7.4.
      Concentration Label Please refer to vial label for lot-specific concentration
      Recommended reaction conditions
      Kinase Assay This protocol is meant to serve as a guideline and may need to be modified for specific applications. Materials Required • Lipid Mixture: 1. Determine the total amount of lipid mixture needed. Each reaction requires 20 µg phosphatidylserine (1.2 µl of 10 mg/ml phosphatidylserine in CHCl3) and 2 µg 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 minutes. 6. Place the lipid mixture in a 40°C water bath for 5 minutes 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&epsilon 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 dH2O; total volume should be 60 µl; prepare a sufficient reaction mixture for all assays to be performed. Protocol 1. Dispense 60 µl Reaction Mixture into each tube and incubate at 30°C. 2. Dilute the PKCε stock solution to 20-50 ng/ml in 10 mM Tris-HCl, pH 7.5, 5 mM DTT, and 0.01% Triton® X-100 detergent. 3. Add 2 µl diluted enzyme to each tube. 4. Incubate for 10 min. 5. Stop the reaction by adding 6 µl 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 3 times with 5 ml of 0.5% phosphoric acid per filter. 9. Transfer membranes to a scintillation vial and count.
      Purity≥85% by SDS-PAGE
      Specific activity≥800 units/mg protein
      Unit definitionOne unit is defined as the amount of enzyme that will transfer 1 nmol phosphate to PKCε Peptide Substrate (Cat. No. 539562) per min at 30°C, pH 7.4.
      Storage Avoid freeze/thaw
      ≤ -70°C
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
      Toxicity Standard Handling
      ReferencesNewton, A.C. 1995. J. Biol. Chem. 270, 28495.
      Dekker, L.V., and Parker, P.J. 1994. Trends Biochem. Sci. 19, 73.
      Nishizuka, Y. 1992. Science 258, 607.
      Bell, R.M., and Burns, D.J. 1991. J. Biol. Chem. 266, 4661.