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324881 PERK, GST-Fusion, Human, Recombinant, E. coli

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324881
  
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      Übersicht

      Replacement Information
      Description
      Overview

      This product has been discontinued.



      Recombinant, human PERK consisting of amino acids 563-1115 fused at the N-terminus to GST and expressed in E. coli. PERK phosphorylates the α-subunit of eukaryotic translation initiation factor 2 (EIF2α), which leads to its inactivation and reduces translation initiation. The active kinase is useful for the study of PERK regulation and for inhibitor screening.

      Note: the calculated MW is 88,000, but the protein runs at ~115,000 on SDS-PAGE.
      Catalogue Number324881
      Brand Family Calbiochem®
      SynonymsEIF2AK3 PKR
      Application Data
      The activity of PERK was measured as outlined in the Recommended Reaction Conditions.
      References
      ReferencesBaltzis, D., et al. 2007. J Biol Chem. 43, 31675.
      Koumenis, C., et al. 2002. Mol Cell Biol. 21, 7405.


      Product Information
      Unit of DefinitionKinase activity is measured as the molar amount of phosphate incorporated into SMAD3 at 30°C using a final concentration of 50 µM ATP.
      FormLiquid
      FormulationIn 50 mM Tris-HCl, 50 mM NaCl, 250 µM DTT, 100 µM EGTA, 100 µM EDTA, 100 µM PMSF, 25% glycerol, pH 7.5.
      Quality LevelMQ100
      Applications
      Biological Information
      Purity≥90% by SDS-PAGE
      Specific ActivityPlease refer to the lot-specific Certificate of Analysis
      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
      Bestellnummer GTIN
      324881 0

      Documentation

      PERK, GST-Fusion, Human, Recombinant, E. coli SDB

      Titel

      Sicherheitsdatenblatt (SDB) 

      PERK, GST-Fusion, Human, Recombinant, E. coli Analysenzertifikate

      TitelChargennummer
      324881

      Literatur

      Übersicht
      Baltzis, D., et al. 2007. J Biol Chem. 43, 31675.
      Koumenis, C., et al. 2002. Mol Cell Biol. 21, 7405.


      Datenblatt

      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.

      Revision06-September-2017 JSW
      SynonymsEIF2AK3 PKR
      Application Data
      The activity of PERK was measured as outlined in the Recommended Reaction Conditions.
      DescriptionRecombinant, human PERK consisting of amino acids 563-1115 fused at the N-terminus to GST and expressed in E. coli. PERK phosphorylates the α-subunit of eukaryotic translation initiation factor 2 (EIF2α), which leads to its inactivation and reduces translation initiation. The active kinase is useful for the study of PERK regulation and for inhibitor screening. Note: the calculated MW is 88,000, but the protein runs at ~115,000 on SDS-PAGE.
      FormLiquid
      FormulationIn 50 mM Tris-HCl, 50 mM NaCl, 250 µM DTT, 100 µM EGTA, 100 µM EDTA, 100 µM PMSF, 25% glycerol, pH 7.5.
      Concentration Label Please refer to vial label for lot-specific concentration
      Recommended reaction conditions
      Kinase assay conditions: Dilute purified PERK in kinase dilution buffer (5 mM MOPS, pH 7.2, 4 mM MgCl2, 2.5 mM β-glycerophosphate, 1 mM EGTA, 400 µM EDTA, 50 ng/µl BSA, 5% glycerol; add DTT to 50 µM just prior to use) to the desired concentration. Mix 10 µl diluted enzyme, 5 µl SMAD3 substrate (200 ng/ml stock), and 5 µl H2O and incubate for 5 min at 30°C. Add 5 µl ATP mixture [(150 µl of 10 mM ATP, 100 µl of 1 mCi/100µl [32P]ATP, and 5.75 ml kinase assay buffer (25 mM MOPS, pH 7.2, 25 mM MgCL2, 12.5 mM β-glycerophosphate, 5 mM EGTA, 2 mM EDTA with 250 µM DTT just prior to use)] and incubate for 15 min at 30°C. Terminate the reaction by spotting 20 µl of the reaction mixture onto a phosphocellulose P81 paper, dry, and wash several times with 1% phosphoric acid prior to counting in the presence of scintillation fluid.
      Purity≥90% by SDS-PAGE
      Specific activityPlease refer to the lot-specific Certificate of Analysis
      Unit definitionKinase activity is measured as the molar amount of phosphate incorporated into SMAD3 at 30°C using a final concentration of 50 µM ATP.
      Storage Avoid freeze/thaw
      ≤ -70°C
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
      Toxicity Standard Handling
      ReferencesBaltzis, D., et al. 2007. J Biol Chem. 43, 31675.
      Koumenis, C., et al. 2002. Mol Cell Biol. 21, 7405.