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342001 Procathepsin K, Human, Recombinant, E. coli

Recombinant, human procathepsin K expressed in E. coli with a methionine residue inserted at amino acid 18 to create a new N-terminal initiation site.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
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342001
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Overview

Replacement Information

Products

Catalog NumberPackaging Qty/Pack
342001-10UG Plastic ampoule 10 μg
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
Catalog 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.