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444208 MMP-1, Proenzyme, Human Rheumatoid Synovial Fibroblast

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

Synonyms: Matrix Metalloproteinase 1, Human Interstitial Collagenase, Collagenase-1

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      Overview

      Replacement Information
      Description
      OverviewNative proMMP-1 from cultured, human rheumatoid synovial fibroblast. Corresponds to the 56 kDa/52 kDa enzyme. May contain some activated enzyme but requires activation by APMA (Cat. No. 164610) for 30 - 60 min. at 30°C just prior to use to obtain fully activated enzyme (46 kDa/42 kDa). May also undergo autocatalysis to yield a 27 kDa/22 kDa active enzyme. Expressed by a large number of cell types. Cleaves fibrillar type I collagen.
      M.W. 56,000/52,000. Note: 1 mU = 1 milliunit.
      Catalogue Number444208
      Brand Family Calbiochem®
      SynonymsMatrix Metalloproteinase 1, Human Interstitial Collagenase, Collagenase-1
      References
      ReferencesLiepinsh, E., et al. 2003. J. Biol. Chem. 278, 25982.
      Pilcher, B.K., et al. 1997. J. Cell Biol. 137, 1445.
      Vallon, R., et al. 1997. Eur. J. Biochem. 244, 81.
      Marcy, A.I., et al. 1991. Biochemistry 30, 6476.
      Stricklin, et al. 1983. Biochemistry 22, 61.
      Product Information
      Unit of DefinitionOne unit is defined as the amount of enzyme that will hydrolyze 1.0 µmol 2,4-DNP-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg-OH per min at 37°C pH 7.0.
      EC number3.4.24.7
      FormLiquid
      FormulationIn 300 mM NaCl, 50 mM Tris-HCl, 5 mM CaCl₂, 1 µM ZnCl₂, 0.05% BRIJ® 35 Detergent, 0.05% NaN₃, pH 7.0.
      Quality LevelMQ100
      Applications
      Biological Information
      Purity≥90% by SDS-PAGE
      SourcePrepared from culture medium of human rheumatoid synovial fibroblasts that has been shown by certified tests to be negative for HBsAg and for antibodies to HIV and HCV.
      Specific Activity≥15 mU/mg protein
      Concentration Label Please refer to vial label for lot-specific concentration
      Physicochemical Information
      Contaminantsno other MMP activity detectable
      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
      Catalogue Number GTIN
      444208 0

      Documentation

      MMP-1, Proenzyme, Human Rheumatoid Synovial Fibroblast SDS

      Title

      Safety Data Sheet (SDS) 

      MMP-1, Proenzyme, Human Rheumatoid Synovial Fibroblast Certificates of Analysis

      TitleLot Number
      444208

      References

      Reference overview
      Liepinsh, E., et al. 2003. J. Biol. Chem. 278, 25982.
      Pilcher, B.K., et al. 1997. J. Cell Biol. 137, 1445.
      Vallon, R., et al. 1997. Eur. J. Biochem. 244, 81.
      Marcy, A.I., et al. 1991. Biochemistry 30, 6476.
      Stricklin, et al. 1983. Biochemistry 22, 61.
      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.

      Revision16-June-2010 RFH
      SynonymsMatrix Metalloproteinase 1, Human Interstitial Collagenase, Collagenase-1
      DescriptionNative proMMP-1 from cultured, human rheumatoid synovial fibroblast. Corresponds to the 56 kDa/52 kDa enzyme. May contain some activated enzyme but requires APMA (Cat. No. 164610; 30-60 minutes at 37°C, just prior to use) to obtain fully activated enzyme (46 kDa/42 kDa). May also undergo autocatalysis to yield a 27 kDa/22 kDa active enzyme. Expressed by a large number of cell types. Cleaves fibrillar Type I collagen. Must be activated just prior to use.
      FormLiquid
      FormulationIn 300 mM NaCl, 50 mM Tris-HCl, 5 mM CaCl₂, 1 µM ZnCl₂, 0.05% BRIJ® 35 Detergent, 0.05% NaN₃, pH 7.0.
      Concentration Label Please refer to vial label for lot-specific concentration
      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 using APMA in DBS. 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.
      SourcePrepared from culture medium of human rheumatoid synovial fibroblasts that has been shown by certified tests to be negative for HBsAg and for antibodies to HIV and HCV.
      EC number3.4.24.7
      Purity≥90% by SDS-PAGE
      Contaminantsno other MMP activity detectable
      Specific activity≥15 mU/mg protein
      Unit definitionOne unit is defined as the amount of enzyme that will hydrolyze 1.0 µmol 2,4-DNP-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg-OH per min at 37°C pH 7.0.
      Storage Avoid freeze/thaw
      ≤ -70°C
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
      Toxicity Standard Handling
      ReferencesLiepinsh, E., et al. 2003. J. Biol. Chem. 278, 25982.
      Pilcher, B.K., et al. 1997. J. Cell Biol. 137, 1445.
      Vallon, R., et al. 1997. Eur. J. Biochem. 244, 81.
      Marcy, A.I., et al. 1991. Biochemistry 30, 6476.
      Stricklin, et al. 1983. Biochemistry 22, 61.

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