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 (ECM). MMP‑16 (MT3-MMP) is found in brain, lung, placenta, smooth muscle cells, and malignant tumor tissues including oral melanoma and renal carcinoma (1). MMP‑16 has been shown to activate proMMP-2 and degrade various ECM components including native collagens (2, 3). MMP‑16 has been proposed to possess the potential to directly enhance the growth and invasiveness of cells in vivo, two critical processes for development and carcinogenesis (4). Structurally, MMP‑16 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 (1). The structure of the catalytic domain in complex with a hydroxamate inhibitor has been solved (5). The recombinant human MMP‑16PC consists of the pro and catalytic domains, which can be activated by treatment with furin.
Recombinant Human MMP-16/MT3-MMP Protein, CF
R&D Systems | Catalog # 1785-MP
Key Product Details
- R&D Systems E. coli-derived Recombinant Human MMP-16/MT3-MMP Protein (1785-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
Accession Number
Structure / Form
Applications
Product Specifications
Source
Ala32-Gly291 (Ile152Asn), with a C-terminal 10-His tag
Purity
Endotoxin Level
N-terminal Sequence Analysis
Predicted Molecular Mass
SDS-PAGE
Activity
The specific activity is >250 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
1785-MP
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl, CaCl2, ZnCl2 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.
|
Background: MMP-16/MT3-MMP
References
- Takino, T. et al. (1995) J. Biol. Chem. 270:23013.
- Shofuda, K. et al. (1997) J. Biol. Chem. 272:9749.
- Shimada, T. et al. (1999) Eur. J. Biochem. 262:907.
- Kang, T. et al. (2000) FASEB J. 14:2559.
- Lang, R. et al. (2004) J. Mol. Biol. 336:213.
Long Name
Alternate Names
Entrez Gene IDs
Gene Symbol
UniProt
Additional MMP-16/MT3-MMP Products
Product Documents for Recombinant Human MMP-16/MT3-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-16/MT3-MMP Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human MMP-16/MT3-MMP Protein, CF
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Protocols
View specific protocols for Recombinant Human MMP-16/MT3-MMP Protein, CF (1785-MP):
- Activation Buffer: 50 mM Tris, 1 mM CaCl2, 0.5% (v/v) Brij-35, pH 9.0
- Assay Buffer: 50 mM Tris, 3 mM CaCl2, 1 µM ZnCl2, pH 8.5
- Recombinant Human MMP‑16/MT3‑MMP (rhMMP-16) (Catalog # 1785-MP)
- Recombinant Human Furin His-tag (rhFurin) (Catalog # 11338-SE)
- Substrate: MCA-Lys-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # ES010)
- Black 96-well Plate
- Plate Reader with Fluorescence Read Capability
- Dilute rhMMP-16 to 80 µg/mL in Activation Buffer.
- Dilute rhFurin to 1.76 µg/mL in Activation Buffer.
- Mix equal volumes of 80 µg/mL of rhMMP-16 and 1.76 µg/mL of rhFurin together.
- Incubate at 37 °C for 1 hour to activate rhMMP-16.
- After incubation, dilute activated rhMMP-16 to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- In plate, load 50 µL of 2 ng/µL rhMMP-16, and start the reaction by adding 50 µL of 20 µM Substrate to wells. As a Substrate Blank, load 50 µL of Assay Buffer and 50 µL of 20 µM Substrate.
- Read at excitation and emission wavelengths of 320 nm and 405 nm, respectively, in kinetic mode for 5 minutes.
- 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-16: 0.1 µg
- Substrate: 10 µM
FAQs for Recombinant Human MMP-16/MT3-MMP Protein, CF
-
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.