324783 EphB4, GST-Fusion, Human, Recombinant, S. frugiperda

324783
  
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      Áttekintés

      Replacement Information
      Description
      Overview

      This product has been discontinued.






      Recombinant, human EPHB4 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. EPHB4 is a member of the tyrosine kinase family and is normally expressed in endothelial and neuronal cells. Ephrin-B ligands undergo phosphorylation on conserved inracellular tyrosines upon interaction with receptors. Aberrant expression of EPHB4 has been reported in prostate cancer and highly malignant breast cancers.
      Catalogue Number324783
      Brand Family Calbiochem®
      SynonymsEPH-related tyrosine kinase receptor B4
      References
      ReferencesKumar, S.R., et al. 2006. Am. J. Pathol. 169, 279.
      Xia, G., et al. 2005. Cancer Res. 65, 4623.
      Sturz, A., et al. 2004. Biochem. Biophys. Res. Commun. 313, 80.
      Berclaz, G., et al. 2003. Ann. Oncol. 14, 220.
      Wang, H.U., et al. 1998. Cell 93, 741.
      Product Information
      Unit of DefinitionOne unit is defined as the amount of enzyme required to transfer 1 nmol phosphate to Poly(Glu,Tyr)<sub>4:1</sub> per min at 30°C, using variable concentrations of ATP (100 nM to 1.71 µM).
      FormLiquid
      FormulationIn 100 mM NaCl, 50 mM Tris-HCl, 5 mM DTT, 20% glycerol, pH 8.0
      Applications
      Biological Information
      Specific Activity≥60 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

      EphB4, GST-Fusion, Human, Recombinant, S. frugiperda Certificates of Analysis

      TitleLot Number
      324783

      References

      Hivatkozások áttekintése
      Kumar, S.R., et al. 2006. Am. J. Pathol. 169, 279.
      Xia, G., et al. 2005. Cancer Res. 65, 4623.
      Sturz, A., et al. 2004. Biochem. Biophys. Res. Commun. 313, 80.
      Berclaz, G., et al. 2003. Ann. Oncol. 14, 220.
      Wang, H.U., et al. 1998. Cell 93, 741.
      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.

      Revision22-August-2007 JSW
      SynonymsEPH-related tyrosine kinase receptor B4
      DescriptionRecombinant, human EPHB4 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. EPHB4 is a member of the tyrosine kinase family and is normally expressed in endothelial and neuronal cells. Ephrin-B ligands undergo phosphorylation on conserved inracellular tyrosines upon interaction with receptors. Aberrant expression of EPHB4 has been reported in prostate cancer and highly malignant breast cancers.
      FormLiquid
      FormulationIn 100 mM NaCl, 50 mM Tris-HCl, 5 mM DTT, 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(Glu,Tyr)4:1, 500 ng/50 µl final concentration EPHB4, GST-Fusion, Human, Recombinant, S. frugiperda, diluted in 10X 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 EPHB4. 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.
      Specific activity≥60 U/mg protein
      Unit definitionOne unit is defined as the amount of enzyme required to transfer 1 nmol phosphate to Poly(Glu,Tyr)4:1 per min at 30°C, using variable concentrations of ATP (100 nM to 1.71 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
      ReferencesKumar, S.R., et al. 2006. Am. J. Pathol. 169, 279.
      Xia, G., et al. 2005. Cancer Res. 65, 4623.
      Sturz, A., et al. 2004. Biochem. Biophys. Res. Commun. 313, 80.
      Berclaz, G., et al. 2003. Ann. Oncol. 14, 220.
      Wang, H.U., et al. 1998. Cell 93, 741.