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444231 MMP-9, Monomer, Human Neutrophil

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444231
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      Description
      OverviewMonomeric, native MMP-9 from stimulated human neutrophils. Requires activation prior to use.
      Note: 1 mU = 1 milliunit.
      Catalogue Number444231
      Brand Family Calbiochem®
      SynonymsMatrix Metalloproteinase 9, Gelatinase B, 92 kDa Type IV Collagenase
      References
      ReferencesReynolds, M. A., et al. 2003. Clin. Chem. 49, 1733.
      Kolkenbrock, H., et al. 1996. Biol. Chem. 377, 529.
      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 number3.4.24.35
      FormLiquid
      FormulationIn 200 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 stimulated neutrophils that have been shown by certified tests to be negative for HBsAg and for antibodies to HIV and HCV.
      Specific Activity≥1000 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

      Documentation

      MMP-9, Monomer, Human Neutrophil SDS

      Title

      Safety Data Sheet (SDS) 

      MMP-9, Monomer, Human Neutrophil Certificates of Analysis

      TitleLot Number
      444231

      References

      Reference overview
      Reynolds, M. A., et al. 2003. Clin. Chem. 49, 1733.
      Kolkenbrock, H., et al. 1996. Biol. Chem. 377, 529.
      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.

      Revision18-July-2007 RFH
      SynonymsMatrix Metalloproteinase 9, Gelatinase B, 92 kDa Type IV Collagenase
      DescriptionMonomeric, native MMP-9 from stimulated human neutrophils. Matrix metalloproteinases (MMP) are members of a family of at least 20 zinc-dependent endoproteinases that function extracellularly at neutral pH. The enzymes are synthesized as latent or inactive zymogens that contain a regulatory propeptide. This propeptide confers latency by binding to the Zn2+ molecule present in the active site. MMP activation can be achieved through autoactivation, via cleavage by other MMPs or proteases, such as furin, urokinase, plasmin, and trypsin, or by treatment with organomercurial comounds. This preparation is highly purified native MMP-9 from stimulated human neutrophils. Requires activation prior to use.
      FormLiquid
      FormulationIn 200 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. 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 stimulated neutrophils that have been shown by certified tests to be negative for HBsAg and for antibodies to HIV and HCV.
      EC number3.4.24.35
      Purity≥90% by SDS-PAGE
      Contaminantsno other MMP activity detectable
      Specific activity≥1000 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 ≤ -70°C
      Avoid freeze/thaw
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
      Toxicity Standard Handling
      ReferencesReynolds, M. A., et al. 2003. Clin. Chem. 49, 1733.
      Kolkenbrock, H., et al. 1996. Biol. Chem. 377, 529.
      Marcy, A.I., et al. 1991. Biochemistry 30, 6476.
      Stricklin, et al. 1983. Biochemistry 22, 61.

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      Categories

      Life Science Research > Proteins and Enzymes > Other Enzymes