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325896 Wee1, GST-Fusion, Human, Recombinant, S. frugiperda

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
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325896
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Overview

Replacement Information

Products

Catalog NumberPackaging Qty/Pack
325896-10UG Plastic ampoule 10 μg
Description
OverviewRecombinant, human Wee1 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. Wee1 is a tyrosine kinase that causes inhibitory phosporylation of Cdk1 and Cdk2 and acts as a mitotic inhibitor. Protects the nucleus from cytoplasmically activated cyclin B1-complexed CDC2 prior to the onset of mitosis.
Catalogue Number325896
Brand Family Calbiochem®
References
ReferencesKatayama, K., et al. 2005. Mol. Cell. Biol. 25, 5725.
Yoshida, T., et al. 2004. Ann. Oncol. 15, 252.
Watanabe, N., et al. 1995. EMBO J. 14, 1878.
Igarashi, M., et al. 1991. Nature 353, 80.
Product Information
Unit of DefinitionOne unit is defined as the amount of enzyme required to transfer 1 nmol phosphate to Poly(Ala,Glu,Lys,Tyr)<sub>6:2:5:1</sub> per min at 30°C, using variable concentrations of ATP (100 nM to 1.55 µM).
EC number2.7.1.112
FormLiquid
FormulationIn 100 mM NaCl, 50 mM Tris-HCl, 5 mM DTT, 4 mM reduced glutathione, 20% glycerol, pH 8.0.
Quality LevelMQ100
Applications
Biological Information
Specific Activity≥1 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
Catalog Number GTIN
325896-10UG 04055977196313

Documentation

Wee1, GST-Fusion, Human, Recombinant, S. frugiperda SDS

Title

Safety Data Sheet (SDS) 

Wee1, GST-Fusion, Human, Recombinant, S. frugiperda Certificates of Analysis

TitleLot Number
325896

References

Reference overview
Katayama, K., et al. 2005. Mol. Cell. Biol. 25, 5725.
Yoshida, T., et al. 2004. Ann. Oncol. 15, 252.
Watanabe, N., et al. 1995. EMBO J. 14, 1878.
Igarashi, M., et al. 1991. Nature 353, 80.

Brochure

Title
Biologics 33.2
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.

Revision10-September-2007 JSW
DescriptionRecombinant, human WEE1 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. WEE1 is a tyrosine kinase that causes inhibitory phosporylation of Cdk1 and Cdk2 and acts as a mitotic inhibitor. Protects the nucleus from cytoplasmically activated cyclin B1-complexed CDC2 prior to the onset of mitosis.
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: Poly(Ala,Glu,Lys,Tyr)6:2:5:1, 1 µg/50 µl final concentration • WEE1, GST-Fusion, Human, Recombinant, S. frugiperda, diluted in 10X Kinase Dilution Buffer to desired concentration (we use 200 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 WEE1. 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 number2.7.1.112
Specific activity≥1 U/mg protein
Unit definitionOne unit is defined as the amount of enzyme required to transfer 1 nmol phosphate to Poly(Ala,Glu,Lys,Tyr)6:2:5:1 per min at 30°C, using variable concentrations of ATP (100 nM to 1.55 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
ReferencesKatayama, K., et al. 2005. Mol. Cell. Biol. 25, 5725.
Yoshida, T., et al. 2004. Ann. Oncol. 15, 252.
Watanabe, N., et al. 1995. EMBO J. 14, 1878.
Igarashi, M., et al. 1991. Nature 353, 80.