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-13 (Collagenase-3) has been demonstrated to degrade a range of extracellular matrix proteins, including collagen types I, II, III, IV, IX, X and XIV, gelatin, aggrecan, perlecan and fibronectin. MMP-13 is distinguished from the other human collagenases by its effecient degradation of type II collagen. MMP-13 is expressed by fibroblasts, chrondrocytes and squamous epithelial cells. Structurally, MMP-13 may be divided into several distinct domains; a pro-domain which is cleaved upon activation; a catalytic domain containing the zinc binding site; a short hinge region and a carboxyl terminal (hemopexin-like) domain.
Recombinant Human MMP-13 Protein, CF
R&D Systems | Catalog # 511-MM
Key Product Details
- R&D Systems NS0-derived Recombinant Human MMP-13 Protein (511-MM)
- 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
Leu20-Cys471
Purity
Endotoxin Level
N-terminal Sequence Analysis
Predicted Molecular Mass
SDS-PAGE
Activity
The specific activity is >2,000 pmol/min/µg, as measured under the described conditions.
Reviewed Applications
Read 2 reviews rated 5 using 511-MM in the following applications:
Formulation, Preparation, and Storage
511-MM
| 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.
|
Background: MMP-13
References
- Jeffery, J.J. (1998) in Collagenase 3. A.J. Barrett, et al. (eds): Handbook of Proteolytic Enzymes, San Diego: Academic Press, p. 1167.
Long Name
Alternate Names
Entrez Gene IDs
Gene Symbol
UniProt
Additional MMP-13 Products
Product Documents for Recombinant Human MMP-13 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-13 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human MMP-13 Protein, CF
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Customer Reviews for Recombinant Human MMP-13 Protein, CF (2)
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Application: In vitro bioactivity in cell cultureVerified Customer | Posted 03/04/2023rhMMP-13 was used as a control to induce inflammation in human THP-1 monocytes differentiated into mature macrophages. Monocytes were treated with lipopolysaccharide (LPS, 1 microgram/mL) and rhMMP-13 (10-100 ng/mL). Results showed a dose-dependent generation of nitrites in these cells, confirming the pro-inflammatory activity of rhMMP-13.
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Application: Immunoassay StandardVerified Customer | Posted 11/10/2017An ELISA was developed to measure MMP-13 in human plasma samples. This recombinant protein was used in this assay to confirm functionality and to build a standard curve.
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Protocols
View specific protocols for Recombinant Human MMP-13 Protein, CF (511-MM):
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% Brij-35 (w/v), pH 7.5 (TCNB)
- Recombinant Human MMP-13 (rhMMP-13) (Catalog # 511-MM)
- p-Aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563) 100 mM stock in DMSO
- Fluorogenic Peptide Substrate I: MCA-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 (Catalog # ES001)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhMMP-13 to 100 µg/mL in Assay Buffer.
- Add APMA to a final concentration of 1 mM.
- Incubate at 37 °C for 2 hours to activate.
- Dilute activated rhMMP-13 to 0.2 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load 50 µL of the 0.2 ng/µL rhMMP-13 into 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.
- 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).
Per Well:
- rhMMP-13: 0.010 µg
- Substrate: 10 µM
FAQs for Recombinant Human MMP-13 Protein, CF
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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: If I use this enzyme at a higher concentration, do I need to change the concentration of APMA to activate it?
A: 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.
-
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: If I use this enzyme at a higher concentration, do I need to change the concentration of APMA to activate it?
A: 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.
-
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: If I use this enzyme at a higher concentration, do I need to change the concentration of APMA to activate it?
A: 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.
-
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: If I use this enzyme at a higher concentration, do I need to change the concentration of APMA to activate it?
A: 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.
-
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.