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475701 MKK6, GST-Fusion, Human, Recombinant, E. coli

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

Synonyms: MEK6, Mitogen Activated Protein Kinase kinase 6, SAPKK3

475701
  
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Replacement Information
Description
Overview

This product has been discontinued.



Recombinant, human MKK6 fused to GST at the N-terminus to GST and expressed in E. coli. MKK6 is a member of the dual specificity protein kinase family, which functions as a mitogen-activated protein (MAP) kinase kinase. It is suitable for labeling MAP kinase substrates.
Catalogue Number475701
Brand Family Calbiochem®
SynonymsMEK6, Mitogen Activated Protein Kinase kinase 6, SAPKK3
Application Data
Increasing amounts of MKK6 were subjected to SDS-PAGE and the purity analyzed by Coomassie Blue staining.

Increasing amounts of MKK6 were used to activate 10 µg p38α/SAPK2a and the activity of p38α/SAPK2a was measured as outlined in the protocols above.
References
ReferencesChabaud-Riou, M. and Firestein, G.S. et al. 2004. Am. J. Pathol. 164, 177.
Zhu, X., et al. 2001. J. Neurochem. 79, 311.
Product Information
Unit of DefinitionOne unit is defined as the amount of enzyme that will activate p38alpha/SAPK2α (0.5 mg/ml) by 1 unit/min using 125 µM ATP at 30°C. One unit of p38α/SAPK2α activity is defined as the amount of enzyme that will transfer 1 pmol phosphate to myelin basic protein per min at 30°C.
FormulationIn 270 mM sucrose, 150 mM NaCl, 50 mM Tris-HCl, 1 mM DTT, 0.1 mM EGTA, pH 7.5.
Quality LevelMQ100
Applications
Biological Information
Purity≥ 90% by SDS PAGE
Specific Activity≥ 50,000 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
Katalogové číslo GTIN
475701 0

Documentation

MKK6, GST-Fusion, Human, Recombinant, E. coli MSDS

Title

Safety Data Sheet (SDS) 

MKK6, GST-Fusion, Human, Recombinant, E. coli Certificates of Analysis

TitleLot Number
475701

References

Přehled odkazů
Chabaud-Riou, M. and Firestein, G.S. et al. 2004. Am. J. Pathol. 164, 177.
Zhu, X., et al. 2001. J. Neurochem. 79, 311.
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.

Revision08-August-2008 RFH
SynonymsMEK6, Mitogen Activated Protein Kinase kinase 6, SAPKK3
Application Data
Increasing amounts of MKK6 were subjected to SDS-PAGE and the purity analyzed by Coomassie Blue staining.

Increasing amounts of MKK6 were used to activate 10 µg p38α/SAPK2a and the activity of p38α/SAPK2a was measured as outlined in the protocols above.
DescriptionRecombinant, human MKK6 fused to GST at the N-terminus and expressed in E. coli. MKK6 is a member of the dual specificity protein kinase family, which functions as a mitogen-activated protein (MAP) kinase kinase. It is suitable for labeling MAP kinase substrates.
FormulationIn 270 mM sucrose, 150 mM NaCl, 50 mM Tris-HCl, 1 mM DTT, 0.1 mM EGTA, pH 7.5.
Concentration Label Please refer to vial label for lot-specific concentration
Recommended reaction conditions
Kinase Activity Assay Activation of p38α/SAPK2a Required Materials •Assay Buffer: 25 mM β-glycerophosphate, 20 mM Tris-HCl, 5 mM EGTA, 1 mM sodium orthovanadate, 1 mM DTT, pH 7.5 • p38&alpha/SPK2a (substrate): in 300 mM NaCl, 50 mM Tris-HCl, 2 mM Dtt, pH 8.5 •MKK6: 100-fold lower concentration than p38α/SAPK2, diluted in Assay Buffer •Magnesium/ATP Mix: 75 mM MgCl2, 500 µM ATP Activation Protocol 1. Determine the reaction volume based on the concentration of p38α/SAPK2a and MKK6. 2. Add 1/8 volume Assay Buffer to a clean microcentrifuge tube on ice. 3. Add 3/8 volume H2O. 4. Add 1/8 volume MKK6. 5. Add 1/8 volume p38α/SAPK2a. 6. Add 1/4 volume Magnesium/ATP Mix. 7. Incubate at 30°C for 1 h. 8. Stop the reaction by placing the tubes on ice. 9. Assay the p38α/SAPK2a activity using the protocol outlined below. Note: The reaction may be dialyzed against 300 mM NaCl, 50 mM Tris-HCl, 2 mM DTT, pH 8.5, diluted with an equal volume of 99% glycerol, and stored at -70°C until use. Activity Assay Required Materials •Assay Buffer: 25 mM β-glycerophosphate, 20 mM Tris-HCl, 5 mM EGTA, 1 mM sodium orthovanadate, 1 mM DTT, pH 7.5 •Substrate: myelin basic protein, 2 mg/ml •p38α/SAPK2a, activated: 2.5-10 ng/µl, activated by MKK6 (above), diluted in Assay Buffer just prior to use •Magnesium/ATP Mix: 75 mM MgCl2, 500 µM ATP •Diluted [γ32P]ATP/Magnesium/ATP Mix: 197 µl Magnesium/ATP Mix, 3 µl (30 Ci) [γ32P]ATP (3000 Ci/mmol) Activity Protocol 1. Add 2.5 µl Assay Buffer to a clean microcentrifuge tube on ice. 2. Add 10 µl Substrate. 3. Add 2.5 µl p38α/SAPK2a (25-100 ng/assay). 4. Add 5 µl Diluted [γ32P]ATP/Magnesium/ATP Mix. 5. Incubate at 30°C for 20 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≥ 90% by SDS PAGE
Specific activity≥ 50,000 units/mg protein
Unit definitionOne unit is defined as the amount of enzyme that will activate p38alpha/SAPK2α (0.5 mg/ml) by 1 unit/min using 125 µM ATP at 30°C. One unit of p38α/SAPK2α activity is defined as the amount of enzyme that will transfer 1 pmol phosphate to myelin basic protein 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
ReferencesChabaud-Riou, M. and Firestein, G.S. et al. 2004. Am. J. Pathol. 164, 177.
Zhu, X., et al. 2001. J. Neurochem. 79, 311.