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

444248
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      Overview

      Replacement Information

      Key Specifications Table

      Pricing & Availability

      Catalog NumberAvailability Packaging Qty/Pack Price Quantity
      444248-10UG
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          Plastic ampoule 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.
          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

          Documentation

          SDS

          Title

          Safety Data Sheet (SDS) 

          Certificates of Analysis

          TitleLot Number
          444248

          References

          Reference overview
          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.

          Brochure

          Title
          Proteases Technical Bulletin
          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.

          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 ≤ -70°C
          Avoid freeze/thaw
          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.