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

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

Synonyms: Protein Kinase A, cAMP Dependent Protein Kinase

539482
  
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Overview

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
Global Trade Item Number
Catalogue Number GTIN
539482 0

Documentation

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

Title

Safety Data Sheet (SDS) 

PKA, His•Tag®, Active, Human, Recombinant, E. coli Certificates of Analysis

TitleLot Number
539482

References

Reference overview
Skalhegg, B.S. and Tasken K., et al. 2000. Front. Biosci. 5, D678.
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.

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 Avoid freeze/thaw
≤ -70°C
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.