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

324881
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      Overview

      Replacement Information

      Pricing & Availability

      Catalog NumberAvailability Packaging Qty/Pack Price Quantity
      324881-5UG
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          Plastic ampoule 5 μg
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          Description
          OverviewRecombinant, 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.
          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

          Documentation

          SDS

          Title

          Safety Data Sheet (SDS) 

          Certificates of Analysis

          TitleLot Number
          324881

          References

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


          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.

          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 ≤ -70°C
          Avoid freeze/thaw
          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.