Recombinant Mouse MMP-3 Protein, CF

R&D Systems | Catalog # 548-MM

R&D Systems
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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse MMP-3 Protein (548-MM)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

P28862

Structure / Form

Pro form

Applications

Enzyme Activity
View all MMP-3 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse MMP-3 protein
Tyr18-Cys477

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Tyr18

Predicted Molecular Mass

52 kDa

SDS-PAGE

60 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The specific activity is >300 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

548-MM
Formulation Supplied as a 0.2 μm filtered solution in Tris, CaCl2, NaCl, Brij-35 and Glycerol.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: MMP-3

Matrix metalloproteinases are a family of zinc and calcium-dependent endopeptidases with the combined ability to degrade all the components of the extracellular matrix. MMP-3 (stromelysin-1) can degrade a broad range of substrates including collagen alpha -chains, aggrecan, laminin, fibronectin, elastin, casein, alpha -1 antitrypsin, myelin basic protein, IL-1 beta, IGFBP-3, pro-MMP-1, pro-MMP-7, pro-MMP-8, pro-MMP-9 and pro-MMP-13. MMP-3 does not cleave the triple helical region of interstitial collagens, a characteristic which distinguishes the stromelysins from the collagenases. The MMP-3 substrate repertoire extends beyond extracellular matrix proteins and implicates MMP-3 in roles other than direct tissue remodeling, for instance, enzyme cascades and cytokine regulation. MMP-3 is expressed by fibroblasts, chrondrocytes, osteoblasts, endothelial cells, smooth muscle cells and macrophages. Structurally, MMP-3 may be divided into several distinct domains, a pro-domain which is cleaved upon activation, a catalytic domain containing the zinc binding site, a short hinge region, and a carboxyl terminal (hemopexin-like) domain.

Long Name

Matrix Metalloproteinase 3

Alternate Names

MMP3, Stromelysin 1

Entrez Gene IDs

4314 (Human); 17392 (Mouse)

Gene Symbol

MMP3

UniProt

P28862

Additional MMP-3 Products

Product Documents for Recombinant Mouse MMP-3 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Mouse MMP-3 Protein, CF

For research use only

Citations for Recombinant Mouse MMP-3 Protein, CF

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Protocols

View specific protocols for Recombinant Mouse MMP-3 Protein, CF (548-MM):

Materials
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Recombinant Mouse (rmMMP-3) (Catalog # 548-MM)
  • p-aminophenylmercuric acetate (APMA) (Sigma, Catalog # A-9563), 100 mM stock in DMSO
  • Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys-(DNP)-NH2 (Catalog # ES002), 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmMMP-3 to 100 µg/mL with 1 mM APMA in Assay Buffer.
  2. To activate, incubate reaction at 37 °C for 1 hour.
  3. Dilute activated rmMMP-3 to 1 ng/µL in Assay Buffer.
  4. Dilute Substrate to 20 µM in Assay Buffer.
  5. In a plate load 50 µL of 1 ng/µL rmMMP-3 to wells, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank of 50 µL Assay Buffer and 50 µL of 20 µM Substrate.
  6. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  7. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

Per Well:

  • mMMP-3: 0.050 µg
  • Substrate: 10 µM

FAQs for Recombinant Mouse MMP-3 Protein, CF

Showing  1 - 4 的 4 FAQs Showing All

    • Q: Can the enzyme be stored after activation, or do I need to use it immediately after activation?
      A: We recommend only activating the amount of enzyme needed for your assay, and recommend activating the enzyme immediately prior to use. Any unactivated enzyme should be stored in aliquots at either the stock concentration at which the enzyme was supplied, or the reconstitution concentration, according to the product datasheet.

    • A: Yes, this enzyme requires activation prior to use.

    • Q: If I use this enzyme at a higher concentration, do I need to change the concentration of APMA to activate it?
      A: We have only optimized activation conditions for one particular concentration of this MMP enzyme as part of our regular QC testing for enzymatic activity. Activating the enzyme at any different concentration would have to be optimized by the end user.

    • A: We supply this enzyme as a mass and calculate its activity relative to mass (pmol/min/µg). We have not calibrated this enzyme to an international standard unit, so we are unable to provide a conversion to units/µg.
Showing  1 - 4 的 4 FAQs Showing All
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