Recombinant Human TACE/ADAM17 Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human 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. 
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human TACE/ADAM17 protein
Arg215-Asn671, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Arg215
Structure / Form
Mature form. Recombinant Human TACE/ADAM17 may be 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

Theoretical MW
52 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
64 kDa, reducing conditions
Publications
Read Publications using
930-ADB 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 reconstitution.
Buffer
Lyophilized from a 0.2 μm filtered solution in Tris, NaCl and Brij-35.
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: 25 mM Tris, 2.5 μM ZnCl2, 0.005% Brij-35 (w/v), pH 9.0 (note: It is extremely important that the assay solution does not contain salt (CaCl2, NaCl, Na2SO4) because it inhibits TACE activity).
  • Recombinant Human TACE/ADAM17 (rhTACE) (Catalog # 930-ADB)
  • 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 rhTACE 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 rhTACE and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 μL Assay Buffer and 50 µL 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:
  • rhTACE: 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 Human 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

    TACE is a member of the ADAM family that contains A Disintegrin And Metalloprotease-like domain. Like other membrane-anchored ADAMs, TACE consists of a pro domain with a cysteine switch and furin cleavage sequence, a catalytic domain with the zinc-binding site and Met-turn expected for reprolysins, a disintegrin-like domain, a cysteine-rich domain, an EGF-like domain, a transmembrane domain, and the cytoplasmic domain. In addition to its ability to release the 17 kDa extracellular form of tumor necrosis factor-alpha (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 Notch-1 receptor. TACE mRNA is present in virtually every tissue and TACE protein resides both on the cell surface and in the cell.

    1. Black, R.A. and J.D. Becherer (1998) in Tumor Necrosis Factor alpha -Converting Enzyme. Barrett, A.J. et al. (eds): Handbook of Proteolytic Enzymes, San Diego: Academic Press, p. 1315.
    2. Primakoff, P. and D.G. Myles (2000) Trends in Genetics 16:83.

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    930-ADB
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    Publications for TACE/ADAM17 (930-ADB)(25)

    We have publications tested in 5 confirmed species: Human, Mouse, Bacteria - E. Coli, Hamster, N/A.

    We have publications tested in 6 applications: Binding Assay, Bioassay, Enzyme Assay, In Vivo, Inhibition, Western Blot.


    Filter By Application
    Binding Assay
    (1)
    Bioassay
    (14)
    Enzyme Assay
    (7)
    In Vivo
    (1)
    Inhibition
    (1)
    Western Blot
    (1)
    All Applications
    Filter By Species
    Human
    (14)
    Mouse
    (4)
    Bacteria - E. Coli
    (1)
    Hamster
    (1)
    N/A
    (5)
    All Species
    Showing Publications 1 - 10 of 25. Show All 25 Publications.
    Publications using 930-ADB Applications Species
    M Maekawa, H Tadaki, D Tomimoto, C Okuma, R Sano, Y Ishii, Y Katsuda, H Yoshiuchi, R Kakefuda, T Ohta, T Sasase A Novel TNF-alpha Converting Enzyme (TACE) Selective Inhibitor JTP-96193 Prevents Insulin Resistance in KK-Ay Type 2 Diabetic Mice and Diabetic Peripheral Neuropathy in Type 1 Diabetic Mice Biol. Pharm. Bull., 2019;42(11):1906-1912. 2019 [PMID: 31685773] (Bioassay, Human) Bioassay Human
    A Bertran, D Khomiak, A Konopka, E Rejmak, E Bulska, J Seco, L Kaczmarek, T Tarragó, R Prades Design and synthesis of selective and blood-brain barrier-permeable hydroxamate-based gelatinase inhibitors Bioorg. Chem., 2019;0(0):103365. 2019 [PMID: 31676116] (Bioassay, Human) Bioassay Human
    A Ikeda, S Shahid, BR Blumberg, M Suzuki, JD Bartlett ADAM10 is Expressed by Ameloblasts, Cleaves the RELT TNF Receptor Extracellular Domain and Facilitates Enamel Development Sci Rep, 2019;9(1):14086. 2019 [PMID: 31575895] (Bioassay, Mouse) Bioassay Mouse
    E Vezzoli, I Caron, F Talpo, D Besusso, P Conforti, E Battaglia, E Sogne, A Falqui, L Petricca, M Verani, P Martufi, A Caricasole, A Bresciani, O Cecchetti, P Rivetti di, G Sancini, O Riess, H Nguyen, L Seipold, P Saftig, G Biella, E Cattaneo, C Zuccato Inhibiting pathologically active ADAM10 rescues synaptic and cognitive decline in Huntington&#039;s disease J. Clin. Invest., 2019;130(0):. 2019 [PMID: 31063986] (Bioassay, Human) Bioassay Human
    Y Ye, J Toczek, K Gona, HY Kim, J Han, M Razavian, R Golestani, J Zhang, TL Wu, M Ghosh, JJ Jung, MM Sadeghi Novel Arginine-containing Macrocyclic MMP Inhibitors: Synthesis, 99mTc-labeling, and Evaluation Sci Rep, 2018;8(1):11647. 2018 [PMID: 30076321] (Enzyme Assay, Human) Enzyme Assay Human
    T Yoneyama, M Gorry, A Sobo-Vujan, Y Lin, L Vujanovic, A Gaither-Da, ML Moss, MA Miller, LG Griffith, DA Lauffenbur, LP Stabile, J Herman, NL Vujanovic ADAM10 Sheddase Activity is a Potential Lung-Cancer Biomarker J Cancer, 2018;9(14):2559-2570. 2018 [PMID: 30026855] (Bioassay, N/A) Bioassay N/A
    JB Schaal, T Maretzky, DQ Tran, PA Tran, P Tongaonkar, CP Blobel, AJ Ouellette, ME Selsted Macrocyclic ?-defensins suppress tumor necrosis factor-? (TNF-?) shedding by inhibition of TNF-? converting enzyme J. Biol. Chem., 2018;0(0):. 2018 [PMID: 29317500] (Bioassay, N/A) Bioassay N/A
    Niacin-mediated Tace activation ameliorates CMT neuropathies with focal hypermyelination EMBO Mol Med, 2016;0(0):. 2016 [PMID: 27799291] (Bioassay, Mouse) Bioassay Mouse
    Mira Jun The Identification of Biochanin A as a Potent and Selective ?-Site App-Cleaving Enzyme 1 (Bace1) Inhibitor Nutrients, 2016;8(10):. 2016 [PMID: 27754406] (Enzyme Assay, N/A) Enzyme Assay N/A
    Mechanistic insights into a TIMP3-sensitive pathway constitutively engaged in the regulation of cerebral hemodynamics Elife, 2016;5(0):. 2016 [PMID: 27476853] (In Vivo, Mouse) In Vivo Mouse
    Show All 25 Publications.

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    Bioinformatics

    Gene Symbol ADAM17
    Uniprot