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 Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
Recombinant Human Aggrecan G1-IGD-G2 (rhAggrecan) Domains (Catalog # 1220-PG)
Recombinant Human ADAMTS1 (rhADAMTS1) (Catalog # 2197-AD)
SDS-PAGE with Silver Staining
Dilute rhAggrecan to 200 µg/mL in Assay Buffer.
Dilute rhADAMTS1 to 56 µg/mL in Assay Buffer.
Mix 25 µL of 200 µg/mL rhAggrecan, 10 µL of 56 µg/mL rhADAMTS1, and 15 µL of Assay Buffer.
Incubate at 37 °C for 24 hours.
Include controls containing equal volumes of 200 µg/mL rhAggrecan and Assay Buffer. Incubate one tube at 37 °C and the other at -20 °C for 24 hours.
Stop the reaction by mixing equal volumes of the incubated samples and reducing gel loading buffer.
Analyze the cleavage by SDS PAGE followed by protein silver staining [or Western blot using anti-human aggrecan antibodies (Catalog # AF1220 and MAB1220)].
rhADAMTS1: 11.2 µg/mL
rhAggrecan: 100 µg/mL
ADAMTS1 (a disintegrin and metalloproteinase with thrombospondin motifs 1), also known as METH1, is the founding member of the family of secreted zinc proteases with a multi-domain structure (1-3). 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. Based on their substrate specificity, ADAMTS1 and associated family members may be key enzymes in degradation of cartilage leading to inflammation and arthritis (4). It is an active protease cleaving alpha -2-macroglobulin (5), aggrecan (6), and versican (7). Compared to ADAMTS4 (aggrecanase 1) and ADAMTS5 (aggrecanase 2), the aggrecanase activity of ADAMTS1 is lower. However, its activity can be enhanced by the binding of cofactor such as fibulin-1 (8). The aggrecanase activity can be inhibited using 5 mM 1,10 Phenanthroline. ADAMTS1 is essential for normal growth and the structure and function of the kidneys, adrenal glands and female reproductive organs (9). It also plays an important role in atherosclerosis (10). It has been shown to inhibit endothelial cell proliferation by direct binding and sequestration of VEGF165 and to inhibit fibroblast migration at high concentrations by binding to FGF-2 (11, 12). The purified recombinant human ADAMTS1 starts at the N‑terminus of the catalytic domain and ends in the beginning of the spacer region.
Vazquez, F. et al. (1999) J. Biol. Chem. 274:23349.
Kuno, K. et al. (1997) J. Biol. Chem. 272:556.
Porter, S. et al. (2005) Biochem. J. 386:15.
Nagase, H. and M. Kashiwagi (2003) Arthritis Res. Ther. 5:94.
Kuno, K. et al. (1999) J. Biol. Chem. 274:18821.
Kuno, K. et al. (2000) FEBS Lett. 478:241.
Russel, D. L. et al. (2003) J. Biol. Chem. 278:42330.
Lee, N., et al. (2005) J. Biol. Chem. 280:34796.
Shindo, T., et al. (2000) J. Clin. Invest. 105:1345.
Krampert, M. et al. (2005) J. Biol. Chem. 280:23844.
A Disintegrin-like and Metalloproteinase Domain with Thrombospondin Motifs 1
Entrez Gene IDs:
a disintegrin-like and metalloprotease (reprolysin type) with thrombospondin type 1 motif, 1; ADAM metallopeptidase with thrombospondin type 1 motif, 1; ADAM-TS 1; ADAMTS1; ADAM-TS1; ADAMTS-1; C3-C5; METH1; METH-1
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
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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