Recombinant Human MMP-15/MT2-MMP Protein, CF

R&D Systems | Catalog # 916-MP

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human MMP-15/MT2-MMP Protein (916-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

NS0

Accession Number

P51511

Applications

Enzyme Activity
View all MMP-15/MT2-MMP Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human MMP-15/MT2-MMP protein
Glu47-Pro565 (Arg128Pro) (Arg129Gly), with a C-terminal 5-His tag

Purity

>70%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

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

N-terminal Sequence Analysis

Glu47

Predicted Molecular Mass

61 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 >200 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

916-MP
Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl and 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-15/MT2-MMP

Matrix metalloproteinases (MMPs) are a family of zinc and calcium dependent endopeptidases with the combined ability to degrade all the components of the extracellular matrix. They play critical roles in tissue remodeling, angiogenesis, tumor invasion, and rheumatoid arthritis (1). MMP-15, also known as MT2-MMP, is a membrane-type MMP that is expressed in many tumor tissues including urothelial carcinoma, oral cancer, ovarian carcinoma, melanoma, and astrocytoma (2). Structurally, MMP-15 consists of the following domains:a pro domain containing a furin cleavage site, a catalytic domain containing the zinc-binding site, a hinge region, a hemopexin-like domain, a transmembrane domain, and a cytoplasmic tail (1). Recombinant Human (rh) MMP-15, consists of the pro domain, catalytic domain, hinge region and hemopexin-like domain. The pro domain contains the mutations R128P and R129G, which prevent activation by furin cleavage. Activation of rhMMP-15 is possible by treatment with rhTrypsin 3 as described in the Activity Assay Protocol.

References

  1. Takino, T. et al. (1995) J. Biol. Chem. 270:23013.
  2. Yana I. and M. Seiki (2004) Handbook of Proteolytic Enzymes (ed. Barrett, et al.) p. 549-551, Academic Press, San Diego.

Long Name

Matrix Metalloproteinase 15/Membrane Type 2 MMP

Alternate Names

MMP15, MT2-MMP

Entrez Gene IDs

4324 (Human)

Gene Symbol

MMP15

UniProt

P51511

Additional MMP-15/MT2-MMP Products

Product Documents for Recombinant Human MMP-15/MT2-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.

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Product Specific Notices for Recombinant Human MMP-15/MT2-MMP Protein, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

For research use only

Related Research Areas

Metalloproteases and Regulators

Citations for Recombinant Human MMP-15/MT2-MMP Protein, CF

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Protocols

View specific protocols for Recombinant Human MMP-15/MT2-MMP Protein, CF (916-MP):

Materials
  • Activation Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 5 µM ZnCl2, 0.05% (w/v) Brij-35, pH 7.5
  • Assay Buffer: 50 mM Tris, 500 mM NaCl, 5 mM CaCl2, 1 µM ZnCl2, 0.02% (w/v) Brij-35, pH 8.0
  • Recombinant Human MMP‑15/MT2‑MMP (rhMMP‑15) (Catalog # 916-MP)
  • Recombinant Human Active Trypsin 3/PRSS3 (rhTrypsin 3) (Catalog # 3714-SE) )
  • 4-(2-Aminoethyl-benzensulfonyl fluoride hydrochloride) (AEBSF) (Tocris, Catalog # 5175), 100 mM stock in deionized water
  • Substrate: MCA-Lys-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # ES010)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Activate rhMMP-15 at 100 µg/mL with 0.1 µg/mL rhTrypsin 3 in Activation Buffer.
  2. Incubate at 37 °C for 2 hours.
  3. Add AEBSF for a final concentration of 1 mM and incubate at room temperature for 15 minutes to stop the reaction.
  4. Dilute activated rhMMP-15 to 1 ng/µL in Assay Buffer.
  5. Dilute Substrate to 20 µM in Assay Buffer.
  6. Load 50 µL of the 1 ng/µL rhMMP-15 in a black well plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate without any rhMMP-15.
  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 (Bachem, Catalog # M-1975).

Per Well:

  • rhMMP-15: 0.05 µg
  • Substrate: 10 µM

FAQs for Recombinant Human MMP-15/MT2-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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