Recombinant Mouse ADAMTS1 Protein, CF

• Reactivity: Mu
• Applications: Enzyme Activity
• Format: Carrier-Free

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 #: P97857
• 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

• Dilutions:
  • Enzyme Activity
• 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

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.

Publications for ADAMTS1 (5867-AD)(4)

Publications using 5867-AD

• SY Cheung, M Sayeed, K Nakuluri, L Li, BJ Feldman MYH9 facilitates autoregulation of adipose tissue depot development JCI Insight, 2021-05-10;6(9):. 2021-05-10 [PMID: 33986190] (Bioassay, Mouse)
• Wong J, Krueger K, Costa M, Aggarwal A, Du H, McLaughlin T, Feldman B A glucocorticoid- and diet-responsive pathway toggles adipocyte precursor cell activity in vivo. Sci Signal, 2016-10-25;9(451):ra103. 2016-10-25 [PMID: 27811141] (Bioassay, Mouse)
• Hellbach N, Weise S, Vezzali R, Wahane S, Heidrich S, Roidl D, Pruszak J, Esser J, Vogel T Neural deletion of Tgfbr2 impairs angiogenesis through an altered secretome. Hum Mol Genet, 2014-07-02;23(23):6177-90. 2014-07-02 [PMID: 24990151] (Bioassay, Human)
• Lee YJ, Koch M, Karl D Variable inhibition of thrombospondin 1 against liver and lung metastases through differential activation of metalloproteinase ADAMTS1. Cancer Res., 2010-01-26;70(3):948-56. 2010-01-26 [PMID: 20103648] (Bioassay, Mouse)

Bioinformatics

• Uniprot:
  • P97857()