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Recombinant Human MMP-14/MT1-MMP (NS0-expressed) Protein, CF

R&D Systems | Catalog # 918-MPN

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human MMP-14/MT1-MMP (NS0-expressed) Protein (918-MPN)
  • 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

P50281

Applications

Enzyme Activity
View all MMP-14/MT1-MMP Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human MMP-14/MT1-MMP protein
Ala21-Ser538, with an Arg108Pro mutation and a C-terminal 6-His tag

Purity

>80%, 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

Ala21

Predicted Molecular Mass

60 kDa

SDS-PAGE

60-65 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 >350 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

918-MPN
Formulation Supplied as a 0.2 μm filtered solution in Sodium Acetate, NaCl, CaCl2, Glycerol.
Shipping The product is shipped with dry ice or equivalent. 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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: MMP-14/MT1-MMP

The first identified 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 1 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 (2). 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, four hemopexin-like domains, a transmembrane domain, and a cytoplasmic tail (3). Recombinant human (rh) MMP-14, consists of the pro domain, catalytic domain, hinge region and hemopexin-like domains. Because it lacks the transmembrane domain, rhMMP-14 is secreted and purified as a soluble protein. The pro domain was engineered to contain an Arg108Pro mutation, which prevents activation by furin cleavage. Activation of rhMMP-14 occurs by treatment with rhTrypsin 3 as described in the Activity Assay Protocol.

References

  1. Seiki, M. (2003) Cancer Lett. 194:1.
  2. Friedl, P. and K. Wolf (2008) Cancer Res. 68:7247.
  3. 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

P50281

Additional MMP-14/MT1-MMP Products

Product Documents for Recombinant Human MMP-14/MT1-MMP (NS0-expressed) 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.

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Product Specific Notices for Recombinant Human MMP-14/MT1-MMP (NS0-expressed) Protein, CF

For research use only

Related Research Areas

Metalloproteases and Regulators

Citations for Recombinant Human MMP-14/MT1-MMP (NS0-expressed) Protein, CF

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Protocols

View specific protocols for Recombinant Human MMP-14/MT1-MMP (NS0-expressed) Protein, CF (918-MPN):

Materials
  • Assay Buffer: 50 mM Tris, 0.15 M NaCl, 10 mM CaCl2, 5 µM ZnCl2, 0.05% (w/v) Brij-35, pH 7.5
  • Recombinant Human MMP‑14/MT1‑MMP (rhMMP-14) (Catalog # 918-MPN)
  • Recombinant Human Trypsin 3 (rhTrypsin 3) (Catalog # 3714-SE)
  • AEBSF (Catalog # 5175), 100 mM stock in deionized water
  • Substrate: MCA-Lys-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # ES010), 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. Activate rhMMP-14 at 100 µg/mL with 0.1 µg/mL rhTrypsin 3 in Assay Buffer.
  2. Incubate at 37 °C for 1 hour.
  3. Stop reaction with 1 mM AEBSF and incubate for 15 minutes at room temperature.
  4. Dilute activated rhMMP-14 to 1 µg/mL in Assay Buffer.
  5. Dilute Substrate to 80 µM in Assay Buffer.
  6. In a plate, load 50 µL of 1 µg/mL rhMMP-14 to wells.  For a Substrate Blank, load 50 µL of Assay Buffer.
  7. Start the reaction by adding 50 µL of 80 µM Substrate to wells.
  8. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  9. 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:

  • rhMMP-14: 0.050 µg
  • Substrate: 40 µM

FAQs for Recombinant Human MMP-14/MT1-MMP (NS0-expressed) 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
View all FAQs for Proteins and Enzymes