>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
<1.0 EU per 1 μg of the protein by the LAL method.
Measured by its ability to cleave the fluorogenic peptide substrate, Abz-TEGEARGSVI-Dap(Dnp)-KK-NH2. The specific activity is >7 pmol/min/μg, as measured under the described conditions. See Activity Assay Protocol on www.RnDSystems.com.
Chinese Hamster Ovary cell line, CHO-derived Phe213-Cys685, with a C-terminal 10-His tag
Recombinant Human ADAMTS4 is prone to proteolytic cleavage at C-terminus. The predominant form of the purified protein lacks the His tag.
Predicted Molecular Mass
58 kDa, reducing conditions
Formulation Supplied as a 0.2 μm filtered solution in Sodium Acetate, CaCl2 and NaCl.
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.
6 months from date of receipt, -20 to -70 °C as supplied.
3 months, -20 to -70 °C under sterile conditions after opening.
Assay Buffer: 50 mM HEPES, 50 mM NaCl, 1 mM CaCl2, 0.05% Brij-35, pH 7.5
Recombinant Human ADAMTS4 (Catalog # 4307-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 rhADAMTS4 to 10 µg/mL in Assay Buffer.
Dilute Substrate to 50 µM in Assay Buffer.
Load 50 µL of 10 µg/mL of rhADAMTS4 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).
rhADAMTS4: 0.5 µg
Substrate: 25 µM
ADAMTS4 (a disintegrin and metalloproteinase with thrombospondin motifs 4), also known as aggrecanase-1, is a member of the family of secreted zinc proteases with a multi-domain structure (1-3). The protein precursors consist of a signal peptide and the following domains: pro, catalytic, disintegrin-like, TS type 1 motif, cysteine-rich, and spacer. It is the only ADAMTS identified that has one TS type I motif. It is an active protease effectively cleaving alpha -2-macroglobulin and aggrecan at multiple sites, and is inhibited by TIMP-3 with inhibition constants in subnanomolar range (4-6). It receives great attention due to the elevation in its mRNA level after treatment with Interleukin-1 (7). However, in a mouse model of osteoarthritis, ADAMTS4 knock-out mice did not exhibit any significant protective effect (8). The purified rhADAMTS4 starts at the catalytic domain and ends before the spacer domain. If desired, the aggrecanase activity can be inhibited by 5 mM 1 10‑phenanthroline and Recombinant Human TIMP‑3 (Catalog # 973-TM).
A Disintegrin-like and Metalloproteinase Domain with Thrombospondin Motifs 4
Entrez Gene IDs:
9507 (Human); 66015 (Rat)
a disintegrin-like and metalloprotease (reprolysin type) with thrombospondintype 1 motif, 4; ADAM metallopeptidase with thrombospondin type 1 motif, 4; ADAM-TS 4; ADAMTS2; ADAMTS-2; ADAMTS4; ADAM-TS4; ADAMTS-4; ADMP-1; ADMP-1EC 126.96.36.199; Aggrecanase 1; aggrecanase-1; EC 3.4.24; KIAA0688A disintegrin and metalloproteinase with thrombospondin motifs 4
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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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