Recombinant Human ADAMTS15 Protein, CF

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

Recombinant Human ADAMTS15 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave Recombinant Human Aggrecan G1-IGD-G2 Domains (Catalog # 1220-PG). Cleavage of 1 µg of rhAggrecan is >90%, as measured under the described conditions.
• Source: Chinese Hamster Ovary cell line, CHO-derived human ADAMTS15 protein
  Gly18-Cys682 (pro) & Phe213-Cys682 (mature), both with a C-terminal 10-His tag
• Accession #: Q8TE58
• N-terminal Sequence: Gly18 & Phe213
• Protein/Peptide Type: Recombinant Enzymes
• Gene: ADAMTS15
• Purity: >80%, 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: 74 kDa (pro), 53 kDa (mature).
  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: 85 kDa and 65 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 MES and NaCl.
• Purity: >80%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Assay Procedure:
  • Dilution Buffer: 50 mM Tris, 0.02% (w/v) Brij-35, pH 7.5
  • 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 Domains (rhAggrecan) (Catalog # 1220-PG)
  • Recombinant Human ADAMTS15 (rhADAMTS15) (Catalog # 5149-AD)
  • SDS-PAGE and/or Western blot
  1. Dilute rhADAMTS15 to 25 μg/mL in Dilution Buffer.
  2. Incubate on ice for 4 hours.
  3. Dilute rhADAMTS15 to 20 µg/mL in Assay Buffer.
  4. Dilute rhAggrecan to 250 µg/mL in Assay Buffer.
  5. Combine equal amounts diluted rhAggrecan and diluted rhADAMTS15. As controls, combine equal amounts Aggrecan and Assay Buffer as well as equal amounts rhADAMTS15 and Assay Buffer.
  6. Incubate the reaction mixture and controls overnight at 37 °C.
  7. Combine 30 µL of reactions (and controls) with 26.2 µL reducing SDS-PAGE sample buffer.
  8. Analyze the cleavage by SDS-PAGE (load 15 µL per lane) followed by protein staining and/or Western blot.
  Per Lane:
  • rhADAMTS15: 0.080 µg
  • rhAggrecan: 1 µg

Notes

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

Alternate Names for Recombinant Human ADAMTS15 Protein, CF

• A disintegrin and metalloproteinase with thrombospondin motifs 15
• a disintegrin-like and metalloprotease (reprolysin type) with thrombospondintype 1 motif, 15
• a disintegrin-like and metalloprotease (reprolysin type) with thrombospondintype 1 motif, 15, preproprotein
• ADAM metallopeptidase with thrombospondin type 1 motif, 15
• ADAM-TS 15
• ADAMTS15
• ADAM-TS15
• ADAMTS-15
• EC 3.4.24
• EC 3.4.24.-
• EC 3.4.24.14
• EC 3.4.24.82
• metalloprotease disintegrin 15 with thrombospondin domains
• MGC126403

Background

ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) is a group of nineteen secreted zinc binding metalloproteases that is involved in development, inflammation, angiogenesis and several pathological conditions (1, 2). They are synthesized as zymogens which have a pro-domain that is removed by furin-like protein convertases. The mature secreted enzymes consist of catalytic, disintegrin-like, type I thrombospondin (TSP) motif, cysteine-rich, and spacer domains followed by zero to fourteen additional TSP motifs. Among ADAMTS proteases, ADAMTS1, -4, -5, -8, and -15 form a subfamily that possess aggrecanase activity (1-3). While ADAMTS4 and -5 (also known as aggrecanse-1 and -2, respectively) (4) have been well studied, the enzymatic activity of ADAMTS15 remains to be explored. It is suggested that ADAMTS15 may be involved in aggrecan degradation and breast cancer (5, 6). ADAMTS15 may also exhibit tumor suppressor activities since it contains genetic mutations in human colorectal carcinomas and restrains tumor growth and invasion in both in vitro and in vivo studies (7). The purified rhADAMTS15 is truncated and ends before the spacer domain.

1. Porter, S. et al. (2005) Biochem. J. 386:15.
2. Jones, G.C. and G.P. Riley (2005) Arthritis Res. Ther. 7:160.
3. Nicholson, A.C. et al. (2005) BMC Evol. Biol. 5:11.
4. Nagase, H. and M. Kashiwagi (2003) Arthritis Res. Ther. 5:94.
5. Porter, S. et al. (2006) Intl. J. Cancer 118:1241.
6. Kevorkian, L. et al. (2004) Arthritis Rheum. 50:131.
7. Viloria, C.G. et al. (2009) Cancer Res. 69:4926.

Bioinformatics

• Gene Symbol: ADAMTS15
• Uniprot:
  • Q8TE58()