Recombinant Human ADAMTS1 Protein, CF

Catalog # Availability Size / Price Qty
2197-AD-020
Product Details
Citations (4)
FAQs
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Recombinant Human ADAMTS1 Protein, CF Summary

Purity
>90%, 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.
Activity
Measured by its ability to cleave Recombinant Human Aggrecan G1-IGD-G2 Domains (Catalog # 1220-PG).
Source
Mouse myeloma cell line, NS0-derived human ADAMTS1 protein
Phe253-Ala734, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
Phe253
Structure / Form
Recombinant Human ADAMTS1 is prone to proteolytic cleavage at C-terminus. The predominant form of the purified protein lacks the His tag.
Predicted Molecular Mass
54 kDa
SDS-PAGE
66 kDa, reducing conditions

Product Datasheets

2197-AD

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 Procedure

Materials
  • 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
  1. Dilute rhAggrecan to 200 µg/mL in Assay Buffer.
  2. Dilute rhADAMTS1 to 56 µg/mL in Assay Buffer.
  3. Mix 25 µL of 200 µg/mL rhAggrecan, 10 µL of 56 µg/mL rhADAMTS1, and 15 µL of Assay Buffer.
  4. Incubate at 37 °C for 24 hours.
  5. 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.
  6. Stop the reaction by mixing equal volumes of the incubated samples and reducing gel loading buffer.
  7. Analyze the cleavage by SDS PAGE followed by protein silver staining [or Western blot using anti-human aggrecan antibodies (Catalog # AF1220 and MAB1220)].
Per Reaction:
  • rhADAMTS1: 11.2 µg/mL
  • rhAggrecan: 100 µg/mL
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Background: ADAMTS1

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.

References
  1. Vazquez, F. et al. (1999) J. Biol. Chem. 274:23349.
  2. Kuno, K. et al. (1997) J. Biol. Chem. 272:556.
  3. Porter, S. et al. (2005) Biochem. J. 386:15.
  4. Nagase, H. and M. Kashiwagi (2003) Arthritis Res. Ther. 5:94.
  5. Kuno, K. et al. (1999) J. Biol. Chem. 274:18821.
  6. Kuno, K. et al. (2000) FEBS Lett. 478:241.
  7. Russel, D. L. et al. (2003) J. Biol. Chem. 278:42330.
  8. Lee, N., et al. (2005) J. Biol. Chem. 280:34796.
  9. Shindo, T., et al. (2000) J. Clin. Invest. 105:1345.
  10. Wight, T.N. (2005) Arterioscler Thromb. Vasc. Biol. 25:12.
  11. Luque, A. et al. (2003) J. Biol. Chem. 278:23656.
  12. Krampert, M. et al. (2005) J. Biol. Chem. 280:23844.
Long Name
A Disintegrin-like and Metalloproteinase Domain with Thrombospondin Motifs 1
Entrez Gene IDs
9510 (Human)
Alternate Names
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

Citations for Recombinant Human ADAMTS1 Protein, CF

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.

4 Citations: Showing 1 - 4
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  1. Cleavage of Fibulin-2 by the aggrecanases ADAMTS-4 and ADAMTS-5 contributes to the tumorigenic potential of breast cancer cells
    Authors: T Fontanil, S ?lvarez-Te, M? Villaronga, Y Mohamedi, L Solares, A Moncada-Pa, JA Vega, O Garc¡a-Su , M P‚rez-Bast, JM Garc¡a-Ped, AJ Obaya, S Cal
    Oncotarget, 2017;0(0):.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  2. ADAMTS-4 promotes neurodegeneration in a mouse model of amyotrophic lateral sclerosis.
    Authors: Lemarchant S, Pomeshchik Y, Kidin I, Karkkainen V, Valonen P, Lehtonen S, Goldsteins G, Malm T, Kanninen K, Koistinaho J
    Mol Neurodegener, 2016;11(1):10.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  3. Matrilin-2 is proteolytically cleaved by ADAMTS-4 and ADAMTS-5.
    Authors: Wang Z, Luo J, Iwamoto S, Chen Q
    Molecules, 2014;19(6):8472-87.
    Species: Primate - Chlorocebus aethiops (African Green Monkey)
    Sample Types: Cell Culture Supernates
    Applications: Bioassay
  4. ADAMTS-1 metalloproteinase promotes tumor development through the induction of a stromal reaction in vivo.
    Authors: Rocks N, Paulissen G, Quesada-Calvo F, Munaut C, Gonzalez ML, Gueders M, Hacha J, Gilles C, Foidart JM, Noel A, Cataldo DD
    Cancer Res., 2008;68(22):9541-50.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay

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