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539513 PKC, Catalytic Subunit, Rat Brain

539513
  
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      Overview

      Replacement Information
      Description
      OverviewNative PKC catalytic subunit purified from rat brain. Prepared by trypsin digestion. Does not require Ca2+ or phosphatidylserine for its activity.

      This product has been discontinued.





      Catalogue Number539513
      Brand Family Calbiochem®
      SynonymsProtein Kinase C-M
      References
      ReferencesVan Renterghem, B., et al. 1994. J. Biol. Chem. 269, 24666.
      Wang, L-Y., et al. 1994. J. Physiol. 475, 431.
      Product Information
      Unit of DefinitionOne unit is defined as the amount of enzyme that will transfer 1.0 nmol of phosphate to histone III-S per min at 30°C.
      FormLiquid
      FormulationIn 100 mM NaCl, 20 mM Tris-HCl, 15 mM DTT, 1 mM EDTA, 1 mM EGTA, 10% glycerol, pH 7.5.
      Applications
      Biological Information
      Purity≥95% by SDS-PAGE
      Specific Activity≥750 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
      539513 0

      Documentation

      PKC, Catalytic Subunit, Rat Brain Certificates of Analysis

      TitleLot Number
      539513

      References

      Reference overview
      Van Renterghem, B., et al. 1994. J. Biol. Chem. 269, 24666.
      Wang, L-Y., et al. 1994. J. Physiol. 475, 431.
      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.

      Revision05-September-2008 RFH
      SynonymsProtein Kinase C-M
      DescriptionNative PKC catalytic subunit purified from rat brain. Prepared by trypsin digestion. Does not require Ca2+ or phosphatidylserine for its activity.
      FormLiquid
      FormulationIn 100 mM NaCl, 20 mM Tris-HCl, 15 mM DTT, 1 mM EDTA, 1 mM EGTA, 10% glycerol, pH 7.5.
      Concentration Label Please refer to vial label for lot-specific concentration
      Recommended reaction conditions
      Reaction Conditions for Protein Kinase C, Catalytic Subunit, Rat Brain

      Assay Solution:
      20 µl 400 mM MES, pH 6.0
      20 µl 10 mM EGTA
      20µl 100 mM MgCl2
      20 µl 5 mg/ml histone III-S (or other substrate)
      40 µl H2O
      20 µl column sample or 20-25 ng purified kinase
      20 µl 1.0 mM ATP (100 µCi/ml)

      Reaction Protocol:
      1. Start the reaction by adding the ATP.
      2. Incubate for 2.5 min at 30°C in an agitating water bath.
      3. Stop the reaction by adding 1 ml of cold 25% trichloroacetic acid.
      4. Add 10 µl of 50 mg/ml BSA per tube.
      5. Vortex for 2 min at 10,000 x g. Aspirate the supernatant.
      6. Repeat steps 3-5 twice using 0.5 ml trichloroacetic acid.
      7. Count dry.

      Terminate the reaction at various time points (e.g. 1, 2, 4, 6, 8 min) to ensure that the reaction is linear.

      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≥750 units/mg protein
      Unit definitionOne unit is defined as the amount of enzyme that will transfer 1.0 nmol of phosphate to histone III-S per min at 30°C.
      Storage Avoid freeze/thaw
      ≤ -70°C
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
      Toxicity Standard Handling
      ReferencesVan Renterghem, B., et al. 1994. J. Biol. Chem. 269, 24666.
      Wang, L-Y., et al. 1994. J. Physiol. 475, 431.