Recombinant Mouse ADAMTS1 Protein, CF

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Product Details

Summary
Reactivity MuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Mouse ADAMTS1 Protein, CF Summary

Details of Functionality
Measured by its ability to cleave Recombinant Human Aggrecan G1-IGD-G2 Domains (Catalog # 1220-PG). rmADAMTS1 achieves >50% substrate cleavage, as measured under the described conditions.
Source
Chinese Hamster Ovary cell line, CHO-derived mouse ADAMTS1 protein
Phe254-Cys725, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Phe254
Structure / Form
Monomer. Recombinant Mouse 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
Purity
>95%, 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
53 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
65-70 kDa, reducing conditions
Publications
Read Publications using
5867-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
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
  • Assay Buffer: 25 mM HEPES, 0.01% (w/v) Brij-35, pH 7.0
  • Recombinant Human Aggrecan Aggrecan G1-IGD-G2 Domains (rhAggrecan) (Catalog # 1220-PG)
  • Recombinant Mouse ADAMTS1 Recombinant Mouse ADAMTS1 (rmADAMTS1) (Catalog # 5867-AD)
  • SDS-PAGE and/or Western blot
  1. Dilute rhAggrecan to 200 µg/mL in Assay Buffer.
  2. Dilute rmADAMTS1 to 20 µg/mL in Assay Buffer.
  3. Combine 25 µL of 200 µg/mL rhAggrecan with 25 µL of 20 µg/mL rmADAMTS1 in a reaction tube.
  4. Incubate the reaction mixture for 24 hours at 37 °C.
  5. For Controls: Combine 25 µL of rhAggrecan solution and 25 µL of Assay Buffer into two tubes. Incubate one tube at 37 °C and the other at -20 °C for 24 hours.
  6. Stop the reaction by adding reduced SDS-PAGE sample buffer.
  7. Analyze the cleavage by SDS PAGE followed by protein staining and/or Western blot.
Per Reaction:
  • rmADAMTS1: 10 µg/mL (0.5 µg)
  • rhAggrecan: 100 µg/mL (5 µg)

Notes

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

Alternate Names for Recombinant Mouse 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 multidomain structure (1-3). The protein precursor consists of a signal peptide and the following domains: pro, catalytic, disintegrinlike, TS type 1 motif, cysteine rich, spacer and a variable number of thrombospondin type 1 motifs. Based on their substrate specificity, ADAMTS1 and associated family members may be key enzymes in the degradation of cartilage leading to inflammation and arthritis (4). It is an active protease cleaving alpha 2macroglobulin (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 a cofactor such as fibulin1 (8). 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 rmADAMTS1 starts at the N terminus of the catalytic domain and ends in the Cys-rich domain.
  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 (5867-AD)(3)

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