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

325896
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      Overview

      Replacement Information

      Pricing & Availability

      Catalog NumberAvailability Packaging Qty/Pack Price Quantity
      325896-10UG
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          Plastic ampoule 10 μg
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          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.
          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

          Documentation

          SDS

          Title

          Safety Data Sheet (SDS) 

          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.
          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 ≤ -70°C
          Avoid freeze/thaw
          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.