Tissue inhibitors of metalloproteinases or TIMPs are a family of proteins that regulate the activation and proteolytic activity of the zinc enzymes known as matrix metalloproteinases (MMPs). There are four members of the family, TIMP-1, TIMP-2, TIMP-3, and TIMP-4. TIMP-2 is a non N-glycosylated protein with a molecular mass of 22 kDa produced by a wide range of cell types, which inhibits MMPs non-covalently by the formation of binary complexes. TIMP-2 also has erythroid‑potentiating and cell growth promoting activities.
Recombinant Human TIMP-2 Protein, CF
R&D Systems | Catalog # 971-TM
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Key Product Details
- R&D Systems CHO-derived Recombinant Human TIMP-2 Protein (971-TM)
- 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
CHO
Accession Number
Applications
Inhibition Activity
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Product Specifications
Source
Chinese Hamster Ovary cell line, CHO-derived human TIMP-2 protein
Cys27-Pro220
Cys27-Pro220
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Cys27
Predicted Molecular Mass
21 kDa
SDS-PAGE
22-23 kDa, reducing conditions
Activity
Measured by its ability to inhibit human MMP-2 cleavage of a fluorogenic peptide substrate Mca-PLGL-Dpa-AR-NH2 (Catalog # ES001).
The IC50 value is approximately 2.2 nM, as measured under the described conditions.
Reviewed Applications
Read 2 reviews rated 4.5 using 971-TM in the following applications:
Formulation, Preparation, and Storage
971-TM
| Formulation | Lyophilized from a 0.2 μm filtered solution in Tris and NaCl. |
| Reconstitution | Reconstitute at 100 μg/mL in sterile 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, and 0.05% Brij-35, pH 7.5. |
| Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Calculators
Background: TIMP-2
Long Name
Tissue Inhibitors of Metalloproteinases 2
Alternate Names
TIMP2
Gene Symbol
TIMP2
UniProt
Additional TIMP-2 Products
Product Documents for Recombinant Human TIMP-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 Human TIMP-2 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human TIMP-2 Protein, CF
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Customer Reviews for Recombinant Human TIMP-2 Protein, CF (2)
4.5 out of 5
2 Customer Ratings
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Application: In vitro bioactivity in cell cultureVerified Customer | Posted 06/09/2021Blocking studies
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Application: Immunoassay StandardVerified Customer | Posted 11/16/2017This recombinant protein was used as an ELISA standard with MAB9711 and BAF971 as matched pair antibodies.
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Protocols
View specific protocols for Recombinant Human TIMP-2 Protein, CF (971-TM):
Materials
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% Brij-35, pH 7.5 (TCNB)
- Recombinant Human TIMP-2 (rhTIMP-2) (Catalog # 971-TM)
- Recombinant Human MMP‑2 (rhMMP‑2) (Catalog # 902-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), 2 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhMMP-2 to 100 µg/mL in Assay Buffer with 1 mM of APMA.
- Incubate at 37 °C for 1 hour.
- Prepare a curve of rhTIMP-2 (MW: 21,500 Da). Make the following serial dilutions in Assay Buffer: 2000, 1000, 500, 300, 200, 150, 100, 50, 25, and 2.5 nM.
- Dilute rhMMP-2 to 12.8 µg/mL in Assay Buffer.
- For each inhibition point mix 25 µL of 12.8 µg/mL rhMMP-2, 16 µL of the rhTIMP-2 curve dilution, and 119 µL of Assay Buffer. Include two enzyme controls of 25 µL of 12.8 µg/mL rhMMP-2 and 135 µL Assay Buffer.
- Incubate reaction mixtures at 37 °C for 2 hours.
- Dilute incubated reaction mixtures by 5 fold in Assay Buffer.
- Dilute Substrate to 10 µM in Assay Buffer.
- In a plate load 50 µL of the 5-fold diluted incubated reaction mixtures to wells.
- Start the reaction by adding 50 µL of 10 µM Substrate to all wells.
Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes. - Derive the 50% inhibiting concentration of rhTIMP-2 (IC50) by plotting RFU/min (or specific activity) vs. concentration with 4-PL fitting.
- The specific activity for rhMMP-2 at each point may be determined using the following formula (if needed):
|
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-2: 0.02 µg
- rhTIMP-2 curve: 20, 10, 5, 3, 2, 1.5, 1.0, 0.5, 0.25, 0.025, and 0 nM
- Substrate: 5 µM