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342001 Procathepsin K, Human, Recombinant, E. coli

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342001
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342001-10UG
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      Description
      OverviewRecombinant, human procathepsin K (amino acids 19-329) (GenBank target symbol = S79895, ACC No. P43235) expressed in E. coli with a methionine residue inserted at amino acid 18 to create a new N-terminal initiation site. Cathepsin K, a member of the papain superfamily of cysteine proteinases, plays an important role in osteoclast-mediated bone resorption and collagen degradation. Cathepsin K is synthesized as an inactive proenzyme that is converted to its mature, active form by proteolytic cleavage of the 99 amino acid propeptide domain. Inhibitors of cathepsin K include leupeptin (Cat. No. 108975) (IC50 = 70 nM), E-64 (Cat. No. 324890) (IC50 = 5 nM), and cystatin (Cat. No. 324891 or 324896). Requires activation prior to use.
      If the activated enzyme is not used immediately, it is recommended to add methyl methanthiosulfonate (1 mM final concentration MMTS).
      Note: 1 mU = 1 milliunit.
      Catalogue Number342001
      Brand Family Calbiochem®
      References
      ReferencesMcQueney, M., et al. 1997. J. Bio. Chem. 272, 13955.
      Bossard, M., et al. 1996. J. Biol. Chem. 271, 12517.
      Drake, F., et al. 1996. J. Biol. Chem. 271, 12511.
      Bromme, D. and Okamoto, K. 1995. Biol. Chem. Hoppe-Seyler 376, 379.
      Baron, R. 1989 Anat. Rec. 224, 317.
      Littlewood-Evans, A.J., et al. 1975. Cancer Res. 57, 5386.
      Nishimura, J.S. et al. 1975. Arch. Biochem. Biophys. 170, 461.
      Product Information
      Activity≥1000 mU/mg protein
      Unit of DefinitionOne unit is defined as the amount of enzyme that will hydrolyze 1 µmole benzyloxycarbonyl-phenylalanine-arginine-7-amido-4-methylcoumarin per min at 37°C, pH 5.5.
      EC number3.4.22.38
      FormLiquid
      FormulationIn 500 mM NaCl, 25 mM Tris, pH 8.0.
      Quality LevelMQ100
      Applications
      Biological Information
      Purity≥95% by SDS-PAGE
      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). Following activation the enzyme is unstable and should include MMTS for storage (see recommended reaction conditions for activation).
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      342001-10UG 04055977194999

      Documentation

      Procathepsin K, Human, Recombinant, E. coli SDS

      Title

      Safety Data Sheet (SDS) 

      Procathepsin K, Human, Recombinant, E. coli Certificates of Analysis

      TitleLot Number
      342001

      References

      Reference overview
      McQueney, M., et al. 1997. J. Bio. Chem. 272, 13955.
      Bossard, M., et al. 1996. J. Biol. Chem. 271, 12517.
      Drake, F., et al. 1996. J. Biol. Chem. 271, 12511.
      Bromme, D. and Okamoto, K. 1995. Biol. Chem. Hoppe-Seyler 376, 379.
      Baron, R. 1989 Anat. Rec. 224, 317.
      Littlewood-Evans, A.J., et al. 1975. Cancer Res. 57, 5386.
      Nishimura, J.S. et al. 1975. Arch. Biochem. Biophys. 170, 461.
      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.

      Revision27-September-2007 RFH
      DescriptionRecombinant, human procathepsin K (amino acids 19-329) (GenBank target symbol = S79895, ACC No. P43235) expressed in E. coli with a methionine residue inserted at amino acid 18 to create a new N-terminal initiation site. Cathepsin K, a member of the papain superfamiliy of cysteine proteinases, plays an important role in osteoclast-mediated bone resorption and collagen degradation. Cathepsin K is synthesized as an inactive proenzyme that is converted to its mature, active form by proteolytic cleavage of the 99 amino acid propeptide domain. Inhibitors of cathepsin K include leupeptin (Cat. No. 108975) (IC50 = 70 nM), E-64 (Cat. No. 324890) (IC50 = 5 nM), and cystatin (Cat. No. 324891 or 324896). Requires activation prior to use.
      FormLiquid
      FormulationIn 500 mM NaCl, 25 mM Tris, pH 8.0.
      Concentration Label Please refer to vial label for lot-specific concentration
      Recommended reaction conditions
      Activation 1. Add an equal volume of 100 mM sodium acetate buffer, 10 mM DTT, 5 mM EDTA, pH 3.9 and incubate at room temperature for 40 min. This will adjust the pH of the procathepsin K to 4.0. 2. If the activated enzyme is not to be used immediately, add methyl methanthiosulfonate (MeS-SO2Me; MMTS) to a final concentration of 1 mM. Aliquot, flash-freeze in liquid nitrogen, and freeze at -70°C. MMTS is a hydrophobic, thiol-reactive compound that modifies cysteines by attaching the uncharged thiomethyl-blocking group to reactive sulhydryl groups. This is a reversible reaction that arrests autoproteolysis. The activity of the enzyme can be restored to (near) unmodified levels by adding an excess of L-cysteine (~3 molar excess over MMTS). Activity Activated cathepsin K activity can be measured in 50 mM MES, 5 mM DTT, 2.5 mM EDTA, pH 5.5, and 100 µM benzyloxycarbonyl-phenylalanine-arginine-7-amido-4-methylcoumarin. Fluorescence is monitored at an excitation wavelength of ~360 nm and an emission wavelength of ~460 nm. To quantitate enzyme activity a 7-amino-4-methylcoumarin standard curve (100-5000 pmol) should be included with each assay. (Note: if MMTS was added to the enzyme after activation, the reaction buffer should also include 3 mM L-cysteine).
      EC number3.4.22.38
      Purity≥95% by SDS-PAGE
      Activity≥1000 mU/mg protein
      Unit definitionOne unit is defined as the amount of enzyme that will hydrolyze 1 µmole benzyloxycarbonyl-phenylalanine-arginine-7-amido-4-methylcoumarin per min at 37°C, pH 5.5.
      Storage Avoid freeze/thaw
      ≤ -70°C
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C). Following activation the enzyme is unstable and should include MMTS for storage (see recommended reaction conditions for activation).
      Toxicity Standard Handling
      ReferencesMcQueney, M., et al. 1997. J. Bio. Chem. 272, 13955.
      Bossard, M., et al. 1996. J. Biol. Chem. 271, 12517.
      Drake, F., et al. 1996. J. Biol. Chem. 271, 12511.
      Bromme, D. and Okamoto, K. 1995. Biol. Chem. Hoppe-Seyler 376, 379.
      Baron, R. 1989 Anat. Rec. 224, 317.
      Littlewood-Evans, A.J., et al. 1975. Cancer Res. 57, 5386.
      Nishimura, J.S. et al. 1975. Arch. Biochem. Biophys. 170, 461.