Recombinant Human ADAMTS1 Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human ADAMTS1 Protein, CF Summary

Details of Functionality
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
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.
Protein/Peptide Type
Recombinant Enzymes
Gene
ADAMTS1
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
54 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
66 kDa, reducing conditions
Publications
Read Publications using
2197-AD in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in Sodium Acetate, CaCl2 and NaCl.
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
  • 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

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human ADAMTS1 Protein, CF

  • 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

Background

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.

  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.

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Publications for ADAMTS1 (2197-AD)(4)

We have publications tested in 3 confirmed species: Human, Mouse, Primate - Chlorocebus aethiops (African Green Monkey).

We have publications tested in 2 applications: Bioassay, Enzyme Assay.


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Bioassay
(3)
Enzyme Assay
(1)
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Human
(2)
Mouse
(1)
Primate - Chlorocebus aethiops (African Green Monkey)
(1)
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Bioinformatics

Gene Symbol ADAMTS1
Uniprot