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171538 AMPK (α1/β1/γ2), His•Tag®, Human, Recombinant, S. frugiperda

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Synonyme: AMP Kinase (α1/β1/γ2)

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      Übersicht

      Replacement Information
      Description
      Overview

      This product has been discontinued.



      Full length, recombinant, human AMPK consisting of subunits α1/β1/γ2, each fused at the C-terminus to a His•Tag® sequence and co-expressed in S. frugiperda insect cells using a baculovirus expression system. AMPK is involved in sensing energy levels in the cell that result from changes in AMP levels. AMPK is an upstream regulator of mTOR activation. The α-subunit contains a kinase domain, the β-subunit contains a glycogen binding domain, and the γ-subunit contains the regulatory AMP binding domain. The active complex is useful in the study of kinase activity regulation and inhibitor screening.
      Catalogue Number171538
      Brand Family Calbiochem®
      SynonymsAMP Kinase (α1/β1/γ2)
      Application Data
      The activity of AMPK (α1/β1/γ2) was measured as outlined in the Recommended Reaction Conditions.
      References
      ReferencesKahn, B.B., et al. 2005. Cell Metab. 1, 15.
      Hardie, G.D., et al. 2004. J. Cell Sci. 117, 5479.
      Product Information
      Unit of DefinitionKinase activity was measured as the molar amount of phosphate incorporated into SAMS peptide substrate (HMRSAMSGLHLVKRR) at 30°C using a final concentration of 50 µM ATP.
      FormLiquid
      FormulationIn 150 mM NaCl, 50 mM Tris-HCl, 250 µM DTT, 100 µM EGTA, 100 µM EDTA, 100 µM PMSF, 25% glycerol, pH 7.5.
      Quality LevelMQ100
      Applications
      Biological Information
      Purity≥70% by SDS-PAGE
      Specific Activity≥897 nmol/min/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
      Bestellnummer GTIN
      171538 0

      Documentation

      AMPK (α1/β1/γ2), His•Tag®, Human, Recombinant, S. frugiperda Analysenzertifikate

      TitelChargennummer
      171538

      Literatur

      Übersicht
      Kahn, B.B., et al. 2005. Cell Metab. 1, 15.
      Hardie, G.D., et al. 2004. J. Cell Sci. 117, 5479.
      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.

      Revision15-September-2008 JSW
      SynonymsAMP Kinase (α1/β1/γ2)
      Application Data
      The activity of AMPK (α1/β1/γ2) was measured as outlined in the Recommended Reaction Conditions.
      DescriptionFull length, recombinant, human AMPK consisting of subunits α1/β1/γ2, each fused at the C-terminus to a His•Tag® sequence and co-expressed in S. frugiperda insect cells using a baculovirus expression system. AMPK is involved in sensing energy levels in the cell that result from changes in AMP levels. AMPK is an upstream regulator of mTOR activation. The α-subunit contains a kinase domain, the β-subunit contains a glycogen binding domain, and the γ-subunit contains the regulatory AMP binding domain. The active complex is useful in the study of kinase activity regulation and inhibitor screening.
      FormLiquid
      FormulationIn 150 mM NaCl, 50 mM Tris-HCl, 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 AMPK in kinase dilution buffer (5 mM MOPS, pH 7.2, 5 mM MgCl2, 2.5 mM β-glycerophosphate, 1 mM EGTA, 400 µM EDTA, 50 ng/µl BSA, 5% glycerol; add DTT to 25 µM just prior to use) to the desired concentration. Mix 10 µl diluted enzyme, 5 µl SAMStide substrate (HMRSAMSGLHLVKRR) (1 mg/ml stock), and 5 µl 500 µM AMP 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≥70% by SDS-PAGE
      Specific activity≥897 nmol/min/mg protein
      Unit definitionKinase activity was measured as the molar amount of phosphate incorporated into SAMS peptide substrate (HMRSAMSGLHLVKRR) 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
      ReferencesKahn, B.B., et al. 2005. Cell Metab. 1, 15.
      Hardie, G.D., et al. 2004. J. Cell Sci. 117, 5479.