Recombinant Mouse TACE/ADAM17 Protein, CF

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

Recombinant Mouse TACE/ADAM17 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave a fluorogenic peptide substrate Mca-PLAQAV-Dpa-RSSSR-NH₂ (Catalog # ES003). The specific activity is >500 pmol/min/µg, as measured under the described conditions. Note: It is extremely important that the assay solution does not contain salt as it inhibits TACE activity.
• Source: Mouse myeloma cell line, NS0-derived mouse TACE/ADAM17 protein
  Pro18-Asp563 and Arg215-Asp563, both with a C-terminal 10-His tag, Pro18-Arg214 and Asp59-Arg214
• Accession #: AAC62934
• N-terminal Sequence: Pro18, Asp59 & Arg215
• Structure / Form: Recombinant Mouse TACE/ADAM17 is prone to proteolytic cleavage at C-terminus. The poly-His tag may not be present in the preparation.
• Protein/Peptide Type: Recombinant Enzymes
• Gene: Adam17
• 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

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 63 kDa, 40 kDa, 23 kDa, 18 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: 76 kDa, 63 kDa, 21 kDa and 18 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 reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in MES and NaCl.
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Reconstitution Instructions: Reconstitute at 100 μg/mL in sterile, deionized water.
• Assay Procedure:
  • Assay Buffer: 50 mM Tris, pH 9.0
  • Recombinant Mouse TACE/ADAM17 (rmTACE) (Catalog # 2978-AD)
  • Substrate: MCA-Pro-Leu-Ala-Gln-Ala-Val-DPA-Arg-Ser-Ser-Ser-Arg-NH₂ (Catalog # ES003) , 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmTACE to 0.2 ng/µL in Assay Buffer.
  2. Dilute Substrate to 20 µM in Assay Buffer.
  3. In a plate, load 50 µL of 0.2 ng/µL rmTACE to wells, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank of 50 µL Assay Buffer and 50 µL of 20 µM Substrate.
  4. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  5. Calculate specific activity:

  Specific Activity (pmol/min/µg) = (Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)) / (amount of enzyme (µg))

  *Adjusted for Substrate Blank
  **Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

  Per Well:
  • rmTACE: 0.01 µg
  • Substrate: 10 µM

Notes

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

Alternate Names for Recombinant Mouse TACE/ADAM17 Protein, CF

• ADAM 17
• ADAM metallopeptidase domain 17
• ADAM metallopeptidase domain 18
• ADAM17
• ADAM18
• CD156b antigen
• CD156b
• CSVP
• disintegrin and metalloproteinase domain-containing protein 17
• EC 3.4.24.86
• MGC71942
• Snake venom-like protease
• TACE
• TACEcSVP
• TNF-alpha convertase
• TNF-alpha converting enzyme
• TNF-alpha-converting enzyme
• tumor necrosis factor, alpha, converting enzyme

Background

Tumor necrosis factor alpha (TNF-alpha ) converting enzyme (TACE) is a member of the ADAM family that contains a disintegrin and metalloprotease-like domain. The domain structure of mouse TACE consists of pro (aa 18-214), catalytic (aa 215-474), disintegrin-like (aa 475-563), cysteine-rich (aa 564-602), Crambin-like (aa 603-671), transmembrane (aa 672-692), and cytoplasmic (aa 693-827) (1, 2). Recombinant mouse TACE ends after the disintegrin-like domain and consists of several forms as described.  In addition to its ability to release the 17 kDa extracellular form of TNF-alpha from the 26 kDa membrane-anchored TNF-alpha , TACE also plays an essential role in shedding ectodomains from a variety of proteins such as L-selectin, transforming growth factor-alpha , amyloid protein precursor, and Notch1 receptor. TACE mRNA is present in virtually every tissue and TACE protein resides both on the cell surface and in the cell (3).

1. Cerretti, D.P. et al. (1999) Cytokine 11:541.
2. Mizui, Y. et al. (1999) Gene 233:67.
3. Black, R.A. (2004) Handbook of Proteolytic Enzymes, Barrett, A.J. et al. eds. pp. 726, Academic Press, San Diego.

Publications for TACE/ADAM17 (2978-AD)(5)

Publications using 2978-AD

• L Scherschin, M Prem, I Kremenetsk, I Tinhofer, P Vajkoczy, AG Karbe, JS Onken Regulation of the Receptor Tyrosine Kinase AXL in Response to Therapy and Its Role in Therapy Resistance in Glioblastoma International Journal of Molecular Sciences, 2022-01-17;23(2):. 2022-01-17 [PMID: 35055167] (Bioassay, Human)
• A-Disintegrin and Metalloproteinase (ADAM) 17 Enzymatically Degrades Interferon-gamma Sci Rep, 2016-08-30;6(0):32259. 2016-08-30 [PMID: 27573075] (Bioassay, Mouse)
• Marczynska J, Ozga A, Wlodarczyk A, Majchrzak-Gorecka M, Kulig P, Banas M, Michalczyk-Wetula D, Majewski P, Hutloff A, Schwarz J, Chalaris A, Scheller J, Rose-John S, Cichy J The role of metalloproteinase ADAM17 in regulating ICOS ligand-mediated humoral immune responses. J Immunol, 2014-08-08;193(6):2753-63. 2014-08-08 [PMID: 25108021] (Bioassay)
• Ramdas V, McBride M, Denby L, Baker A Canonical transforming growth factor-beta signaling regulates disintegrin metalloprotease expression in experimental renal fibrosis via miR-29. Am J Pathol, 2013-10-06;183(6):1885-96. 2013-10-06 [PMID: 24103556] (Bioassay, Rat)
• Wang Y, Huang Z, Nayak P, Matthews B, Warburton D, Shi W, Sanchez-Esteban J Strain-induced differentiation of fetal type II epithelial cells is mediated via the integrin alpha6beta1-ADAM17/tumor necrosis factor-alpha-converting enzyme (TACE) signaling pathway. J Biol Chem, 2013-07-25;288(35):25646-57. 2013-07-25 [PMID: 23888051] (Bioassay, Mouse)

Bioinformatics

• Gene Symbol: Adam17
• Uniprot:
  • AAC62934()