Recombinant Human MMP-17 Protein, CF Summary
Product Specifications
Ala39-Ser531 (Arg125Pro) & (Ala182Thr), with a C-terminal 6-His tag
Analysis
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
7796-MP
| Formulation | Supplied as a 0.2 μm filtered solution in MES, 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.
|
Assay Procedure
- Activation Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB) + 100 µM ZnCl2
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, pH 7.5
- Recombinant Human MMP-17 (rhMMP-17) (Catalog # 7796-MP)
- p-aminophenylmercuric acetate (APMA) (Sigma, Catalog # A-9563), 100 mM stock in DMSO
- Substrate: MCA-PLAQAV-Dpa-RSSSR-NH2 (Catalog # ES003), 2 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: Gemini EM by Molecular Devices) or equivalent
- Activate rhMMP-17 at 100 µg/mL with 1 mM APMA in Activation Buffer.
- Incubate at room temperature for 2 hours.
- Dilute activated rhMMP-17 to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load 50 µL of the 2 ng/µL rhMMP-17 into 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‑17.
- Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), 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 (Bachem, Catalog # M-1975).
- rhMMP-17: 0.1 µg
- Substrate: 10 µM
Reconstitution Calculator
Background: MMP-17
Matrix metalloproteinase 17 (MMP‑17) is a zinc metalloproteinase that is an integral membrane protein (1). It belongs to the membrane‑type MMP (MT‑MMP) subfamily, and is also known as MT4‑MMP. It is associated with the cell membrane by a glycosylphosphatidylinositol (GPI) anchor (1). It has the potential to function as a pro‑tumor necrosis factor‑a (TNF‑a) convertase (2). Unlike MMP‑14 (MT1‑MMP), it is a poor activator of pro‑MMP‑2. MMP‑17 is relatively poor at digesting components of the extracellular matrix, but does cleave fibrinogen and fibrin (2). MMP‑17 is known to activate the aggrecanase ADAMTS‑4 through proteolysis (3). MMP‑17 is expressed in many tissues with the highest levels seen in the brain, colon, ovary, testis, and kidney papilla (4). It has been detected in several human cancers, including gliomas, prostate carcinomas, and breast carcinomas (5). Recombinant Human MMP‑17 was expressed without its GPI anchoring sequence, resulting in its secretion as a soluble protein.
- Itoh, Y. et al. (1999) J. Biol. Chem. 274:34260.
- English, W. R. et al. (2000) J. Biol. Chem. 275:14046.
- Gao, G. et al. (2004) J. Biol. Chem. 279:10042.
- Srichai, M. B. et al. (2011) PLoS ONE 6:e17099.
- Sohail, A. et al. (2008) Cancer Metastasis Rev. 27:289.
Product Specific Notices
Coomassie is a registered trademark of Imperial Chemical Industries Ltd.Citations for Recombinant Human MMP-17 Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 2
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Discovery of potent and specific inhibitors targeting the active site of MMP-9 from the engineered SPINK2 library
Authors: H Yano, D Nishimiya, Y Kawaguchi, M Tamura, R Hashimoto
PLoS ONE, 2020-12-29;15(12):e0244656.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Kallikrein-Related Peptidase 14 Activates Zymogens of Membrane Type Matrix Metalloproteinases (MT-MMPs)-A CleavEx Based Analysis
Authors: K Falkowski, E Bielecka, IB Thøgersen, O Boche?ska, K P?aza, M Kali?ska, L S?siadek, M Magoch, A P?cak, M Wi?niewska, N Gruba, M Wysocka, A Wojtysiak, M Brzezi?ska, K Sychowska, A Pejkovska, M Rehders, G Butler, CM Overall, K Brix, G Dubin, A Lesner, A Kozik, JJ Enghild, J Potempa, T Kantyka
Int J Mol Sci, 2020-06-19;21(12):.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay
FAQs
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Can the enzyme be stored after activation, or do I need to use it immediately after activation?
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.
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If I use this enzyme at a higher concentration, do I need to change the concentration of APMA to activate it?
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.
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Does this MMP enzyme need to be activated to work?
Yes, this enzyme requires activation prior to use.
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What is the activity of this enzyme in units/µg?
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.
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