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Recombinant Human MMP-14/MT1-MMP Protein, CF

R&D Systems | Catalog # 918-MP

R&D Systems
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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human MMP-14/MT1-MMP Protein (918-MP)
  • 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

E. coli

Accession Number

Structure / Form

Pro and catalytic domains

Applications

Enzyme Activity
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Product Specifications

Source

E. coli-derived human MMP-14/MT1-MMP protein
Ala21-Gly284, with a C-terminal 10-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

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

N-terminal Sequence Analysis

Ala21

Predicted Molecular Mass

31 kDa

SDS-PAGE

31 kDa, reducing conditions

Activity

Measured by its ability to cleave a fluorogenic peptide substrate Mca-KPLGL-Dpa-AR-NH2 (Catalog # ES010).
The specific activity is >450 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

918-MP
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, CaCl2 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-14/MT1-MMP

As the first member of membrane type (MT) MMPs, MMP-14, also known as MT1-MMP, plays an important role in extracellular matrix (ECM) remodeling by being able to degrade type I collagen, activate pro-MMP-2 and process cell adhesion molecules such as CD44 and integrin alpha V (1). MMP-14 is therefore a key enzyme in many physiological and pathological processes such as angiogenesis and tumor invasion. Structurally, MMP-14 consists of the following domains: a pro domain containing the furin cleavage site, a catalytic domain containing the zinc-binding site, a hinge region, a hemopexin-like domain, a transmembrane domain, and a cytoplamasic tail (2). Recombinant Human MMP-14 consists of the pro and catalytic domains, which can be activated by treatment with furin as described in the Activity Assay Protocol.

References

  1. Seike, M. (2003) Cancer Lett. 194:1.
  2. Sato, H. et al. (1994) Nature 370:61.

Long Name

Matrix Metalloproteinase 14/Membrane Type 1 MMP

Alternate Names

MMP-X1, MMP14, MT-MMP1, MT1-MMP

Entrez Gene IDs

4323 (Human)

Gene Symbol

MMP14

UniProt

Additional MMP-14/MT1-MMP Products

Product Documents for Recombinant Human MMP-14/MT1-MMP Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Human MMP-14/MT1-MMP Protein, CF

For research use only

Related Research Areas

Citations for Recombinant Human MMP-14/MT1-MMP Protein, CF

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Protocols

View specific protocols for Recombinant Human MMP-14/MT1-MMP Protein, CF (918-MP):

Materials
  • Activation Buffer: 50 mM Tris, 1 mM CaCl2, 0.5% (w/v) Brij-35, pH 9.0
  • Assay Buffer: 50 mM Tris, 3 mM CaCl2, 1 µM ZnCl2, pH 8.5
  • Recombinant Human MMP‑14/MT1‑MMP (rhMMP-14) (Catalog # 918-MP)
  • Recombinant Human Furin His-tag (rhFurin) (Catalog # 11338-SE
  • Substrate: MCA-Lys-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # ES010), 2 mM stock in DMSO
  • Black 96-well Plate
  • Plate Reader with Fluorescence Read Capability
  1. Activate rhMMP-14 at 80 µg/mL with 1.72 µg/mL rhFurin in Activation Buffer.
  2. Incubate at 37 °C for 1.5 hours.
  3. Dilute activated rhMMP-14 to 1 ng/µL in Assay Buffer.
  4. Dilute Substrate to 80 µM in Assay Buffer.
  5. In a plate load 50 µL of 1 ng/µL rhMMP-14 to wells, and for a Substrate Blank load 50 µL of Assay Buffer.
  6. Start the reaction by adding 50 µL of 80 µM Substrate to wells.
  7. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
  8. 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.

Per Well:

  • rhMMP-14: 0.050 µg
  • Substrate: 40 µM
  • FAQs for Recombinant Human MMP-14/MT1-MMP Protein, CF

    Showing  1 - 3 of 3 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.

    • Q: Does this MMP enzyme need to be activated to work?

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

    • Q: What is the activity of this enzyme in units/µg?

      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.

    • 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.

    • Q: Does this MMP enzyme need to be activated to work?

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

    • Q: What is the activity of this enzyme in units/µg?

      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.

    • 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.

    • Q: Does this MMP enzyme need to be activated to work?

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

    • Q: What is the activity of this enzyme in units/µg?

      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 - 3 of 3 FAQs Showing All
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