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325889 p21 Activated Kinase 1, GST-Fusion, Human, Recombinant, S. frugiperda

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

Synonyms: PAK1

325889
  
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Přehled

Replacement Information
Description
OverviewRecombinant, human PAK1 fused at the N-terminus to a GST-His6-thrombin cleavage site sequence and expressed in S. frugiperda insect cells using a baculovirus expression system. PAK1 is a serine/threonine kinase that interacts with GTP-bound but not GDP-bound CDC42/p21 and RAC1. Phosphorylation at Thr423 by PDK activates PAK1. It is known as an important modulator of cytoskeletal reorganization and GTPase effector that links the Rho-related GTPases to the JNK MAP kinase pathway.
Catalogue Number325889
Brand Family Calbiochem®
SynonymsPAK1
References
ReferencesSingh, R.R., et al. 2005. J. Biol. Chem. 280 18130.
Chen, S., et al. 2003. J. Biol. Chem. 278, 20029.
Sells, M.A., et al. 1997. Curr. Biol. 7, 202.
Product Information
Unit of DefinitionOne unit is defined as the amount of enzyme required to transfer 1 nmol phosphate to tetra(LRRWSLG) per min at 30°C using variable concentrations from of ATP (0.1-27.1 µM).
EC number2.7.1.37
FormLiquid
FormulationIn 100 mM NaCl, 50 mM Tris-HCl, 5 mM DTT, 4 mM reduced glutathione, 20% glycerol, pH 8.0.
Applications
Biological Information
Specific Activity≥325 U/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
Katalogové číslo GTIN
325889 0

Documentation

p21 Activated Kinase 1, GST-Fusion, Human, Recombinant, S. frugiperda MSDS

Title

Safety Data Sheet (SDS) 

p21 Activated Kinase 1, GST-Fusion, Human, Recombinant, S. frugiperda Certificates of Analysis

TitleLot Number
325889

References

Přehled odkazů
Singh, R.R., et al. 2005. J. Biol. Chem. 280 18130.
Chen, S., et al. 2003. J. Biol. Chem. 278, 20029.
Sells, M.A., et al. 1997. Curr. Biol. 7, 202.
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.

Revision23-August-2007 JSW
SynonymsPAK1
DescriptionRecombinant, human PAK1 fused at the N-terminus to a GST-His6-thrombin cleavage site sequence and expressed in S. frugiperda insect cells using a baculovirus expression system. PAK1 is a mamalian signaling kinase that interacts with GTP-bound but not GDP-bound CDC42/P21 and RAC1. It is known as an important modulator of cytoskeletal reorganization and GTPase effector that links the Rho-related GTPases to the JNK MAP kinase pathway.
FormLiquid
FormulationIn 100 mM NaCl, 50 mM Tris-HCl, 5 mM DTT, 4 mM reduced glutathione, 20% glycerol, pH 8.0.
Concentration Label Please refer to vial label for lot-specific concentration
Recommended reaction conditions
96-Well Membrane Binding Kinase Activity Assay Protocol:
Materials Required • Standard Kinase Assay Buffer: 125 mM HEPES-NaOH, 7.5 mM MgCl2, 7.5 mM MnCl2, 7.5 µM sodium -orthovanadate, 2.5 mM DTT, pH 7.5 • 10X Kinase Dilution Buffer: 500 mM HEPES-NaOH, 2.5 mg/ml PEG20.000, 10 mM DTT, pH 7.5 • Substrate: tetra(LRRWSLG), 5 µg/50 µl final concentration • PAK1, GST-Fusion, Human, Recombinant, S. frugiperda, diluted in 1X Kinase Dilution Buffer to desired concentration (we use 25 ng in a 50 µl reaction) • 33P-γ-ATP • ATP Mix: 1 µM ATP in H2O with 1 x 106 cpm/5 µl • 96-well V-bottom MTP (polypropylene) plates (Nunc Cat. No. 442587) • 96-well Multiscreen Vacuum Manifold (Millipore Cat. No. MAVM096OR) • 96-well Multiscreen Filterplates, phosphocellulose membrane (Millipore Cat. No. MSPH N6B50) and glass fiber (Millipore Cat. No. MSFC N6B50) • 2% H3PO4 • 0.9% NaCl
Activity Assay Protocol 1. In a polypropylene V-bottom plate add 20 µl Kinase Assay Buffer 2. If desired, add 5 µl test substance (e.g., inhibitor, activator, etc.; in 10% DMSO) 3. Add 10 µl Substrate (diluted in H2O). 4. Add 10 µl diluted PAK1. 5. Add 5 µl ATP Mix. 6. Mix on a shaker and incubate at 30°C for 80 min. 7. Stop the reaction with 50 µl H3PO4. 8. Prewet the filter plate with 200 µl H2O. 9. Filter the reaction mix by applying vacuum. 10. Wash 3 times with 200 µl 0.9% NaCl. 11. Add 200 µl scintillation fluid to each well and count on a scintillation counter.
EC number2.7.1.37
Specific activity≥325 U/mg protein
Unit definitionOne unit is defined as the amount of enzyme required to transfer 1 nmol phosphate to tetra(LRRWSLG) per min at 30°C using variable concentrations from of ATP (0.1-27.1 µM).
Storage Avoid freeze/thaw
≤ -70°C
Do Not Freeze Ok to freeze
Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
Toxicity Standard Handling
ReferencesSingh, R.R., et al. 2005. J. Biol. Chem. 280 18130.
Chen, S., et al. 2003. J. Biol. Chem. 278, 20029.
Sells, M.A., et al. 1997. Curr. Biol. 7, 202.