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444248 MMP-13, Proenzyme, His•Tag®, Human, Recombinant, S. frugiperda

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444248
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444248-10UG
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      Ampoule plast. 10 μg
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      Description
      OverviewFull-length recombinant, human MMP-13 fused to a His•Tag® sequence and expressed in S. frugiperda insect cells. This 452 amino acid proenzyme contains the N-terminal propeptide, the Ca2+- and Zn2+-binding catalytic domain, the hinge region, and the C-terminal hemopexin domain. Hydrolyzes collagen type II 5 to 6 times faster than collagens Type I and III. Exhibits high activity towards gelatin and degrades α1-antichymotrypsin and plasminogen activator inhibitor-2 (PAI-2).
      Note: 1 mU = 1 milliunit.
      Catalogue Number444248
      Brand Family Calbiochem®
      SynonymsCollagenase-3
      References
      ReferencesHerget, J., et al. 2003. Am. J. Physiol. 285, 4199.
      Nagase, H. 1997. Biol. Chem. 378, 151.
      Knäuper, V., et al. 1996. J. Biol. Chem. 271, 1544.
      Knauper, V., et al. 1996. J. Biol. Chem. 271, 17124.
      Pei, D., and Weiss, S.J. 1996. J. Biol. Chem. 271, 9135.
      Knight, C.G., et al. 1992. FEBS Lett. 296, 263.
      Product Information
      Unit of DefinitionOne unit is defined as the amount of APMA-activated enzyme that will hydrolyze 1.0 µmol MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH₂ (<a href = "/Products/ProductDisplay.asp→catNO=03-32-5032">Cat. No. 03-32-5032</a>) per min at 37°C, pH 7.5.
      FormLiquid
      FormulationIn 150 mM NaCl, 50 mM Tris-HCl, 5 mM CaCl₂, pH 7.5.
      Quality LevelMQ100
      Applications
      Biological Information
      Purity≥90% by SDS-PAGE
      Specific Activity≥50 mU/mg protein
      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
      Référence GTIN
      444248-10UG 04055977186482

      Documentation

      MMP-13, Proenzyme, His•Tag®, Human, Recombinant, S. frugiperda FDS

      Titre

      Fiche de données de sécurité des matériaux (FDS) 

      MMP-13, Proenzyme, His•Tag®, Human, Recombinant, S. frugiperda Certificats d'analyse

      TitreNuméro de lot
      444248

      Références bibliographiques

      Aperçu de la référence bibliographique
      Herget, J., et al. 2003. Am. J. Physiol. 285, 4199.
      Nagase, H. 1997. Biol. Chem. 378, 151.
      Knäuper, V., et al. 1996. J. Biol. Chem. 271, 1544.
      Knauper, V., et al. 1996. J. Biol. Chem. 271, 17124.
      Pei, D., and Weiss, S.J. 1996. J. Biol. Chem. 271, 9135.
      Knight, C.G., et al. 1992. FEBS Lett. 296, 263.
      Fiche technique

      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.

      Revision18-July-2007 RFH
      SynonymsCollagenase-3
      DescriptionFull-length recombinant, human MMP-13 fused to a His•Tag® sequence and expressed in S. frugiperda insect cells. This 452 amino acid proenzyme contains an N-terminal propeptide which confers latency to the proenzyme, a Ca2+- and Zn2+- binding catalytic domain, a hinge region, and a C-terminal hemopexin domain. Hydrolyzes collagen type II 5-6 times faster than collagens type I and III. Exhibits high activity towards gelatin and degrades α1-antichymotrypsin and plasminogen activator inhibitor-2.
      FormLiquid
      FormulationIn 150 mM NaCl, 50 mM Tris-HCl, 5 mM CaCl₂, pH 7.5.
      Recommended reaction conditions

      Organomercurial Activation Protocol This protocol is provided only as a general guide. Researchers should standardize this assay for their own specific needs and should consult published literature. The following protocol is from Stricklin, et al., which describes the use of p-aminophenylmercuric acetate (APMA) to activate pro-MMP. This protocol is also adaptable to other types of organomercurals, such as p-(hydroxymercuric) benzoate (PHMB), phenylmercuric chloride (PMC), or mersalyl. 1. Prepare a 10-50 mM stock solution of APMA (or other organomercurial compound) in 0.1 M NaOH just prior to use. Although not absolutely necessary, the stock solution may be adjusted to pH 11 with 5 N HCl (see Marcy, A.I., et al.). 2. To initiate the activation mix the proenzyme solution with the APMA solution at a 10:1 volume ratio (MMP:APMA). If a higher concentration of APMA is desired, increase the concentration of the stock solution. Do not exceed the 10:1 ratio, as this could result in significant changes in pH. 3. Incubate the mixture at 37°C for 2-3 h. It is recommended that an analytical run be conducted first to determine the optimal incubation time. For example, a small-scale experiment with a fixed concentration of pro-MMP and organomercurial would be incubated as described above. Remove aliquots of the sample at various time points during the incubation. Stop the reaction by the addition of SDS-PAGE sample buffer (e.g., 10 µl 2X sample buffer to 10 µl aliquot) and heat the samples to 95°C. The progress of activation can be monitored qualitatively by analyzing the aliquots on a 12% SDS-PAGE gel. 4. The activated MMP can be used without removing the APMA from the mixture. Please refer to Marcy, A.I., et al. for removal of organomercurials by gel filtration.
      Purity≥90% by SDS-PAGE
      Specific activity≥50 mU/mg protein
      Unit definitionOne unit is defined as the amount of APMA-activated enzyme that will hydrolyze 1.0 µmol MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH₂ (Cat. No. 03-32-5032) per min at 37°C, pH 7.5.
      Storage Avoid freeze/thaw
      ≤ -70°C
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
      Toxicity Standard Handling
      ReferencesHerget, J., et al. 2003. Am. J. Physiol. 285, 4199.
      Nagase, H. 1997. Biol. Chem. 378, 151.
      Knäuper, V., et al. 1996. J. Biol. Chem. 271, 1544.
      Knauper, V., et al. 1996. J. Biol. Chem. 271, 17124.
      Pei, D., and Weiss, S.J. 1996. J. Biol. Chem. 271, 9135.
      Knight, C.G., et al. 1992. FEBS Lett. 296, 263.