Recombinant Human MMP-10 Protein, CF Summary
Product Specifications
Tyr18-Cys476
Analysis
Product Datasheets
910-MP
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.
910-MP
| Formulation | Supplied as a 0.2 μm filtered solution in MES, NaCl, CaCl2 and Brij-35. |
| 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
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
- Recombinant Human MMP-10 (rhMMP-10) (Catalog # 910-MP)
- p-Aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563) 100 mM stock in DMSO
- Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: Gemini EM by Molecular Devices) or equivalent
- Activate rhMMP-10 at 100 µg/mL in Assay Buffer containing 1 mM APMA.
- Incubate at 37 °C for 2 hours.
- Dilute activated rhMMP-10 to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load 50 µL of the 2 ng/µL rhMMP-10 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-10.
- 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-10: 0.1 µg
- Substrate: 10 µM
Background: MMP-10
Matrix metalloproteinases are a family of zinc and calcium dependent endopeptidases with the combined ability to degrade all the components of the extracellular matrix. MMP-10 (stromelysin 2) degrades a broad range of substrates including gelatin, collagen types III, IV and V, fibronectin, aggrecan, and pig cartilage proteoglycan. MMP-10 can activate other MMPs such as MMP-1 and MMP-8. MMP-10 is expressed in keratinocytes, T cells, menstrual endometrium and a few tumor samples. Structurally, MMP-10 may be divided into four distinct domains: a pro-domain which is cleaved upon activation, a catalytic domain containing the zinc binding site; a short linker region, and a carboxyl terminal hemopexin-like domain.
Citations for Recombinant Human MMP-10 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 - 4
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Functional human iPSC-derived alveolar-like cells cultured in a miniaturized 96?Transwell air-liquid interface model
Authors: T Bluhmki, S Traub, AK Müller, S Bitzer, E Schruf, MT Bammert, M Leist, F Gantner, JP Garnett, R Heilker
Scientific Reports, 2021-08-23;11(1):17028.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Matrix metalloproteinase-10 protects against acute kidney injury by augmenting epidermal growth factor receptor signaling
Authors: C Hu, Y Zuo, Q Ren, X Sun, S Zhou, J Liao, X Hong, J Miao, L Zhou, Y Liu
Cell Death & Disease, 2021-01-12;12(1):70.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Biodistribution of Nanostructured Lipid Carriers in Mice Atherosclerotic Model
Authors: L Devel, G Almer, C Cabella, F Beau, M Bernes, P Oliva, F Navarro, R Prassl, H Mangge, I Texier
Molecules, 2019-09-26;24(19):.
Species: Human
Sample Types: Peptide
Applications: Bioassay -
Simple pseudo-dipeptides with a P2' glutamate: a novel inhibitor family of matrix metalloproteases and other metzincins.
Authors: Devel L, Beau F, Amoura M, Vera L, Cassar-Lajeunesse E, Garcia S, Czarny B, Stura E, Dive V
J Biol Chem, 2012-06-11;287(32):26647-56.
Applications: Enzyme Assay
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 ratio of pro- and active forms of this enzyme prior to activation?
We do not measure the ratio of pro- to active MMP-10 prior to activation. We measure the activity of each lot after activation to ensure it passes our activity specification.
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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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