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539482 PKA, His•Tag®, Active, Human, Recombinant, E. coli

539482
  
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      Aperçu

      Replacement Information
      Description
      Overview

      This product has been discontinued.



      Recombinant, human, catalytic subunit of PKA fused to at the N-terminus to a His•Tag® sequence and expressed in E.coli. PKA regulates a number of cellular functions including gene expression, carbohydrate metabolism, Ca2+ homeostasis, and ion channel function. Suitable for labeling PKA substrates.

      Catalogue Number539482
      Brand Family Calbiochem®
      SynonymsProtein Kinase A, cAMP Dependent Protein Kinase
      Application Data
      Increasing amounts of PKA were subjected to SDS-PAGE and purity analyzed by Coomassie Blue staining.

      Increasing amounts of PKA were analyzed for activity as outlined in the protocol above.
      References
      ReferencesSkalhegg, B.S. and Tasken K., et al. 2000. Front. Biosci. 5, D678.
      Product Information
      Unit of DefinitionOne unit is defined as the amount of enzyme required to transfer 1 pmol phosphate to PKA peptide (GRTGRRNSI) per min at 30°C.
      FormLiquid
      FormulationIn 150 mM KCl, 30 mM K₂HPO₄, 1 mM DTT, 1 mM EDTA, 50% glycerol, pH 7.4.
      Quality LevelMQ100
      Applications
      Biological Information
      Purity≥95% by SDS PAGE
      Specific Activity≥ 500,000 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

      PKA, His•Tag®, Active, Human, Recombinant, E. coli Certificats d'analyse

      TitreNuméro de lot
      539482

      Références bibliographiques

      Aperçu de la référence bibliographique
      Skalhegg, B.S. and Tasken K., et al. 2000. Front. Biosci. 5, D678.
      Fiche technique

      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.

      Revision04-September-2008 RFH
      SynonymsProtein Kinase A, cAMP Dependent Protein Kinase
      Application Data
      Increasing amounts of PKA were subjected to SDS-PAGE and purity analyzed by Coomassie Blue staining.

      Increasing amounts of PKA were analyzed for activity as outlined in the protocol above.
      DescriptionRecombinant, human, catalytic subunit of PKA fused to at the N-terminus to a His•Tag® sequence and expressed in E. coli. PKA regulates a number of cellular functions including gene expression, carbohydrate metabolism, Ca2+ homeostasis, and ion channel function. Suitable for labeling PKA substrates.
      FormLiquid
      FormulationIn 150 mM KCl, 30 mM K₂HPO₄, 1 mM DTT, 1 mM EDTA, 50% glycerol, pH 7.4.
      Concentration Label Please refer to vial label for lot-specific concentration
      Recommended reaction conditions
      Kinase Activity Assay Required Materials •Assay Buffer: 25 mM β-glycerophosphate, 20 mM Tris-HCl, 5 mM EGTA, 1 mM DTT, pH 7.5 •Substrate: 500 µM GRTGRRNSI peptide substrate •PKA: 2.5-10 ng/µl PKA diluted in Assay Buffer just prior to use •Magnesium/ATP Mix: 75 mM MgCl2, 500 µM ATP •Diluted [γ-32P]ATP/Magnesium/ATP Mix: Mix 197 µl Magnesium/ATP Mix with 3 µl (30 µCi) [γ-32P]ATP (3000 Ci/mmol) Protocol 1. Pipette 10 µl Assay Buffer into a clean microcentrifuge tube on ice. 2. Add 10 µl Substrate. 3. Add 10 µl diluted PKA. 4. Add 10 µl Diluted [γ-32P]ATP/Magnesium/ATP Mix. 5. Incubate at 30°C; for 10 min. 6. Stop the reaction by placing the tubes on ice. 7. Spot 10 µl of each reaction on P81 paper and allow to bind for 30 s. 8. Wash by immersing the paper in 0.75% phosphoric acid and gently shaking. Repeat for a total of 3 washes. 9. Wash as in step 8 using acetone. 10. Dry using an infrared light. 11. Determine the cpm using a scintillation counter or imager.
      Purity≥95% by SDS PAGE
      Specific activity≥ 500,000 units/mg protein
      Unit definitionOne unit is defined as the amount of enzyme required to transfer 1 pmol phosphate to PKA peptide (GRTGRRNSI) per min at 30°C.
      Storage ≤ -70°C
      Avoid freeze/thaw
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
      Toxicity Standard Handling
      ReferencesSkalhegg, B.S. and Tasken K., et al. 2000. Front. Biosci. 5, D678.