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-2 (gelatinase A), a type IV collagenase, can degrade a broad range of substrates including type IV, V, VII and X collagens as well as elastin and fibronectin. It is believed to act synergistically with interstitial collagenase (MMP-1) in the degradation of fibrillar collagens as it degrades their denatured gelatin forms. MMP-2 has been shown to be associated with many connective tissue cells as well as neutrophils, macrophages and monocytes. Structurally, MMP-2 may be divided into several distinct domains: a pro-domain which is cleaved upon activation; a catalytic domain containing the zinc binding site; a fibronectin-like domain thought to play a role in substrate targeting; and a carboxyl terminal (hemopexin-like) domain containing 2 N-linked glycosylation sites. The amino acid sequences of the proenzymes are identical between mouse and rat.
Recombinant Mouse/Rat MMP-2 Protein, CF
R&D Systems | Catalog # 924-MP
Key Product Details
- R&D Systems NS0-derived Recombinant Mouse/Rat MMP-2 Protein (924-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
Ile34-Cys662
Purity
Endotoxin Level
N-terminal Sequence Analysis
Predicted Molecular Mass
SDS-PAGE
Activity
The specific activity is >1,500 pmol/min/μg, as measured under the described conditions.
Reviewed Applications
Read 1 review rated 5 using 924-MP in the following applications:
Formulation, Preparation, and Storage
924-MP
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, CaCl2, NaCl, Brij-35 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.
|
Background: MMP-2
Long Name
Alternate Names
Gene Symbol
UniProt
Additional MMP-2 Products
Product Documents for Recombinant Mouse/Rat MMP-2 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 Mouse/Rat MMP-2 Protein, CF
For research use only
Citations for Recombinant Mouse/Rat MMP-2 Protein, CF
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Customer Images
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Application: Zymogram Positive controlVerified Customer | Posted 04/06/2018The Zymogram shows two concentrations of Recombinant MMP-2 versus MMP-9. They are being used as positive controls for the sample in lane 10 of a bone shaft taken from a female Sprague-Dawley rat.We will be using this for our Zymography positive controls and for publications in the future!
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Protocols
View specific protocols for Recombinant Mouse/Rat MMP-2 Protein, CF (924-MP):
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
- Recombinant Mouse/Rat MMP-2 (rm/rMMP-2) (Catalog # 924-MP)
- p-aminophenylmercuric acetate (APMA) (Sigma, Catalog # A-9563), 100 mM Stock in DMSO
- Substrate: 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 rm/rMMP-2 to 40 µg/mL with 1 mM APMA in Assay Buffer.
- Incubate at 37 °C for 2 hours to activate rm/rMMP-2.
- Dilute activated rm/rMMP-2 to 0.2 ng/µL in Assay Buffer.
- Dilute Substrate to 80 µM in Assay Buffer.
- Load 50 µL of 0.2 ng/µL rm/rMMP-2 in a plate, and start the reaction by adding 50 µL of 80 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 80 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:
- rm/rMMP-2: 0.010 µg
- Substrate: 40 µM
FAQs for Recombinant Mouse/Rat MMP-2 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.
