ADAMTS5 (a disintegrin and metalloproteinase with thrombospondin motifs 5), also known as aggrecanase-2 and ADAMTS11, is a member of the family of secreted zinc proteases with a multi-domain structure (1, 2). The protein precursors consist of signal peptide and following domains: pro, catalytic, disintegrin-like, TS type 1 motif, cysteine-rich, spacer and a variable number of TS type 1 motifs. ADAMTS5 is an active protease effectively cleaving alpha 2-macroglobulin (3), aggrecan (4), and brevican (5), and is inhibited by TIMP-3 with inhibition constants in the subnanomolar range (6). Based on the murine model studies (7, 8), this protease may be a key enzyme in the degradation of cartilage leading to osteoarthritis and rheumatoid arthritis. The purified rhADAMTS5 starts at the N-terminus of the catalytic domain and ends at the C-terminus of the TSP-1 domain. The amino acid sequence of rhADAMTS5 is 98%, 97%, and 96% identical to that of canine, bovine, and mouse/rat. The aggrecanase activity can be inhibited by 5 mM 1,10-phenanthroline and recombinant human TIMP-3 (Catalog # http://www.rndsystems.com/product_results.aspx?k=973-TM">973-TM).
Recombinant Human ADAMTS5 Protein, CF
R&D Systems | Catalog # 2198-AD
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Key Product Details
- R&D Systems NS0-derived Recombinant Human ADAMTS5 Protein (2198-AD)
- 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
NS0
Accession Number
Structure / Form
Recombinant Human ADAMTS5 is prone to proteolytic cleavage at C-terminus. The predominant form of the purified protein lacks the His tag.
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human ADAMTS5 protein
Ser262-Pro622, with a C-terminal 10-His tag
Ser262-Pro622, with a C-terminal 10-His tag
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Ser262
Predicted Molecular Mass
41 kDa
SDS-PAGE
46-51 kDa, reducing conditions
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Abz-TEGEARGSVI-Dap(Dnp)-KK-NH2.
The specific activity is >5 pmol/min/μg, as measured under the described conditions.
The specific activity is >5 pmol/min/μg, as measured under the described conditions.
Reviewed Applications
Read 1 review rated 4 using 2198-AD in the following applications:
Formulation, Preparation, and Storage
2198-AD
| Formulation | Lyophilized from a 0.2 μm filtered solution in Sodium Acetate, CaCl2 and NaCl. |
| Reconstitution | Reconstitute at 200 μg/mL in sterile, deionized water.
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| 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.
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Calculators
Background: ADAMTS5
References
- Porter, S. et al. (2005) Biochem. J. 386:15.
- Nagase, H. and M. Kashiwagi (2003) Arthritis Res. Ther. 5:94.
- Tortorella, M.D. et al. (2004) J. Biol. Chem. 279:17554.
- Vankemmelbeke, M.N. et al. (2001) Eur. J. Biochem. 268:1259.
- Nakada, M. et al. (2005) Acta Neurophathol (Berl). 110:239.
- Kashiwagi, M. et al. (2001) J. Biol. Chem. 276:12501.
- Glasson, S.S. et al. (2005) Nature, 434: 644.
- Stanton, H. et al. (2005) Nature, 434:648.
Long Name
A Disintegrin-like and Metalloproteinase Domain with Thrombospondin Motifs 5
Alternate Names
ADAMTS11, ADMP-2, Aggrecanase 2
Gene Symbol
ADAMTS5
UniProt
Additional ADAMTS5 Products
Product Documents for Recombinant Human ADAMTS5 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 ADAMTS5 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human ADAMTS5 Protein, CF
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Protocols
View specific protocols for Recombinant Human ADAMTS5 Protein, CF (2198-AD):
Materials
- Assay Buffer: 50 mM Tris, 100 mM NaCl, 5 mM CaCl2, 0.05% Brij-35, pH 7.5
- Recombinant Human ADAMTS5 (rhADAMTS5) (Catalog # 2198-AD)
- Substrate: WAAG-3R (Anaspec, Catalog # 60431-1), 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 rhADAMTS5 to 10 µg/mL in Assay Buffer.
- Dilute Substrate to 50 µM in Assay Buffer.
- Load 50 µL of 10 µg/mL of rhADAMTS5 into a plate, and start the reaction by adding 50 µL of 50 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
- Read at excitation and emission wavelengths of 340 nm and 420 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 Abz-Gly-OH (Bachem, Catalog # E-2920).
- rhADAMTS5: 0.5 µg
- Substrate: 25 µM
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