Recombinant Human TACE/ADAM17 Protein, CF

Catalog # Availability Size / Price Qty
930-ADB-010

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Recombinant Human TACE/ADAM17 Protein, CF Summary

Product Specifications

Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
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
Analysis
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.
Predicted Molecular Mass
52 kDa
SDS-PAGE
64 kDa, reducing conditions

Product Datasheets

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930-ADB

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

930-ADB

Formulation Lyophilized from a 0.2 μm filtered solution in Tris, NaCl and Brij-35.
Reconstitution Reconstitute at 100 μg/mL in sterile, deionized water.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Assay Procedure

Materials
  • 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
  • Reconstitution Calculator

    Reconstitution Calculator

    The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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    Background: TACE/ADAM17

    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.

    References
    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.
    Long Name
    TNF-alpha Converting Enzyme
    Entrez Gene IDs
    6868 (Human); 11491 (Mouse)
    Alternate Names
    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

    Citations for Recombinant Human TACE/ADAM17 Protein, CF

    R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

    29 Citations: Showing 1 - 10
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    1. Alpha-1 Antitrypsin Reduces Disease Progression in a Mouse Model of Charcot-Marie-Tooth Type 1A: A Role for Decreased Inflammation and ADAM-17 Inhibition
      Authors: N Zhukovsky, M Silvano, T Filloux, S Gonzalez, KH Krause
      International Journal of Molecular Sciences, 2022-07-03;23(13):.
      Species: Human
      Sample Types: Recombinant Protein
      Applications: Bioassay
    2. Oncogenic KRAS promotes growth of lung cancer cells expressing SLC3A2-NRG1 fusion via ADAM17-mediated shedding of NRG1
      Authors: DH Shin, SH Kim, M Choi, YK Bae, C Han, BK Choi, SS Kim, JY Han
      Oncogene, 2021-11-06;0(0):.
      Species: Human
      Sample Types: Recombinant Proteins
      Applications: Bioassay
    3. ADAM17 promotes the invasion of hepatocellular carcinoma via upregulation MMP21
      Authors: Y Xiang, L Liu, Y Wang, B Li, J Peng, D Feng
      Cancer Cell Int, 2020-10-21;20(0):516.
      Species: Human
      Sample Types: Whole Cells
      Applications: Bioassay
    4. TIMP-3 facilitates binding of target metalloproteinases to the endocytic receptor LRP-1 and promotes scavenging of MMP-1
      Authors: AP Carreca, VM Pravatà, M Markham, S Bonelli, G Murphy, H Nagase, L Troeberg, SD Scilabra
      Sci Rep, 2020-07-21;10(1):12067.
      Species: Human
      Sample Types: Recombinant Protein
      Applications: Surface Plasmon Resonance
    5. Design and synthesis of selective and blood-brain barrier-permeable hydroxamate-based gelatinase inhibitors
      Authors: A Bertran, D Khomiak, A Konopka, E Rejmak, E Bulska, J Seco, L Kaczmarek, T Tarragó, R Prades
      Bioorg. Chem., 2019-10-18;0(0):103365.
      Species: Human
      Sample Types: Protein
      Applications: Bioassay
    6. ADAM10 is Expressed by Ameloblasts, Cleaves the RELT TNF Receptor Extracellular Domain and Facilitates Enamel Development
      Authors: A Ikeda, S Shahid, BR Blumberg, M Suzuki, JD Bartlett
      Sci Rep, 2019-10-01;9(1):14086.
      Species: Mouse
      Sample Types: Protein
      Applications: Bioassay
    7. Inhibiting pathologically active ADAM10 rescues synaptic and cognitive decline in Huntington's disease
      Authors: 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
      J. Clin. Invest., 2019-05-06;130(0):.
      Species: Human
      Sample Types: Recombinant Protein
      Applications: Bioassay
    8. 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
      Authors: M Maekawa, H Tadaki, D Tomimoto, C Okuma, R Sano, Y Ishii, Y Katsuda, H Yoshiuchi, R Kakefuda, T Ohta, T Sasase
      Biol. Pharm. Bull., 2019-01-01;42(11):1906-1912.
      Species: Human
      Sample Types: Protein
      Applications: Bioassay
    9. Novel Arginine-containing Macrocyclic MMP Inhibitors: Synthesis, 99mTc-labeling, and Evaluation
      Authors: Y Ye, J Toczek, K Gona, HY Kim, J Han, M Razavian, R Golestani, J Zhang, TL Wu, M Ghosh, JJ Jung, MM Sadeghi
      Sci Rep, 2018-08-03;8(1):11647.
      Species: Human
      Sample Types: Peptide
      Applications: Enzyme Assay
    10. ADAM10 Sheddase Activity is a Potential Lung-Cancer Biomarker
      Authors: 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
      J Cancer, 2018-06-23;9(14):2559-2570.
      Applications: Bioassay
    11. Macrocyclic ?-defensins suppress tumor necrosis factor-? (TNF-?) shedding by inhibition of TNF-? converting enzyme
      Authors: JB Schaal, T Maretzky, DQ Tran, PA Tran, P Tongaonkar, CP Blobel, AJ Ouellette, ME Selsted
      J. Biol. Chem., 2018-01-09;0(0):.
      Applications: Bioassay
    12. Niacin-mediated Tace activation ameliorates CMT neuropathies with focal hypermyelination
      EMBO Mol Med, 2016-12-01;0(0):.
      Species: Mouse
      Sample Types: Whole Cells
      Applications: Bioassay
    13. The Identification of Biochanin A as a Potent and Selective ?-Site App-Cleaving Enzyme 1 (Bace1) Inhibitor
      Authors: Mira Jun
      Nutrients, 2016-10-14;8(10):.
      Applications: Enzyme Assay
    14. Mechanistic insights into a TIMP3-sensitive pathway constitutively engaged in the regulation of cerebral hemodynamics
      Elife, 2016-08-01;5(0):.
      Species: Mouse
      Sample Types: In Vivo
      Applications: In Vivo
    15. Fibulin-1 Binds to Fibroblast Growth Factor 8 with High Affinity: Effects on Embryo Survival
      J Biol Chem, 2016-07-08;0(0):.
      Species: Human
      Sample Types: Protein
      Applications: Bioassay
    16. ADAM8 as a drug target in pancreatic cancer.
      Authors: Schlomann U, Koller G, Conrad C, Ferdous T, Golfi P, Garcia A, Hofling S, Parsons M, Costa P, Soper R, Bossard M, Hagemann T, Roshani R, Sewald N, Ketchem R, Moss M, Rasmussen F, Miller M, Lauffenburger D, Tuveson D, Nimsky C, Bartsch J
      Nat Commun, 2015-01-28;6(0):6175.
      Species: Human
      Sample Types: Whole Cells
      Applications: Bioassay
    17. Matrix metalloproteinase-8 plays a pivotal role in neuroinflammation by modulating TNF-alpha activation.
      Authors: Lee E, Han J, Woo M, Shin J, Park E, Kang J, Moon P, Baek M, Son W, Ko Y, Choi J, Kim H
      J Immunol, 2014-07-21;193(5):2384-93.
      Applications: Enzyme Assay
    18. ADAM17-mediated shedding of FcgammaRIIIA on human NK cells: identification of the cleavage site and relationship with activation.
      Authors: Lajoie L, Congy-Jolivet N, Bolzec A, Gouilleux-Gruart V, Sicard E, Sung H, Peiretti F, Moreau T, Vie H, Clemenceau B, Thibault G
      J Immunol, 2013-12-13;192(2):741-51.
      Species: Human
      Sample Types: Recombinant Protein
      Applications: Bioassay
    19. A new paradigm for enzymatic control of alpha-cleavage and beta-cleavage of the prion protein.
      Authors: McDonald A, Dibble J, Evans E, Millhauser G
      J Biol Chem, 2013-11-18;289(2):803-13.
      Species: Bacteria - E. Coli
      Sample Types: Cell Lysates
      Applications: Enzyme Assay
    20. Striking reduction of amyloid plaque burden in an Alzheimer's mouse model after chronic administration of carmustine.
      Authors: Hayes C, Dey D, Palavicini J, Wang H, Patkar K, Minond D, Nefzi A, Lakshmana M
      BMC Med, 2013-03-26;11(0):81.
      Species: Hamster
      Sample Types: Whole Cells
      Applications: Inhibition
    21. Unlike for human monocytes after LPS activation, release of TNF-α by THP-1 cells is produced by a TACE catalytically different from constitutive TACE.
      Authors: Moreira-Tabaka H, Peluso J, Vonesch JL, Hentsch D, Kessler P, Reimund JM, Dumont S, Muller CD
      PLoS ONE, 2012-03-30;7(3):e34184.
      Species: Human
      Sample Types: Whole Cells
      Applications: Enzyme Assay
    22. A disintegrin and metalloenzyme (ADAM) 17 activation is regulated by α5β1 integrin in kidney mesangial cells.
      Authors: Gooz P, Dang Y, Higashiyama S, Twal WO, Haycraft CJ, Gooz M
      PLoS ONE, 2012-03-08;7(3):e33350.
      Species: Human
      Sample Types: Fluorogenic Peptide Substrate, Protein
      Applications: Binding Assay, Enzyme Assay
    23. Regulated release and functional modulation of junctional adhesion molecule A by disintegrin metalloproteinases.
      Authors: Koenen RR, Pruessmeyer J, Soehnlein O, Fraemohs L, Zernecke A, Schwarz N, Reiss K, Sarabi A, Lindbom L, Hackeng TM, Weber C, Ludwig A
      Blood, 2009-03-03;113(19):4799-809.
      Species: Human
      Sample Types: Recombinant Protein, Whole Cells
      Applications: Bioassay, Enzyme Assay
    24. The ADAM10 prodomain is a specific inhibitor of ADAM10 proteolytic activity and inhibits cellular shedding events.
      Authors: Moss ML, Bomar M, Liu Q, Sage H, Dempsey P, Lenhart PM, Gillispie PA, Stoeck A, Wildeboer D, Bartsch JW, Palmisano R, Zhou P
      J. Biol. Chem., 2007-09-25;282(49):35712-21.
      Species: Human
      Sample Types: Recombinant Protein
      Applications: Bioassay
    25. Microparticles of human atherosclerotic plaques enhance the shedding of the tumor necrosis factor-alpha converting enzyme/ADAM17 substrates, tumor necrosis factor and tumor necrosis factor receptor-1.
      Authors: Canault M, Leroyer AS, Peiretti F, Leseche G, Tedgui A, Bonardo B, Alessi MC, Boulanger CM, Nalbone G
      Am. J. Pathol., 2007-09-14;171(5):1713-23.
      Species: Human
      Sample Types: Peptide
      Applications: Bioassay
    26. The shedding activity of ADAM17 is sequestered in lipid rafts.
      Authors: Tellier E, Rebsomen L
      Exp. Cell Res., 2006-09-05;312(20):3969-80.
      Species: Human
      Sample Types: Whole Cells
      Applications: Bioassay
    27. Nardilysin enhances ectodomain shedding of heparin-binding epidermal growth factor-like growth factor through activation of tumor necrosis factor-alpha-converting enzyme.
      Authors: Nishi E, Hiraoka Y, Yoshida K, Okawa K, Kita T
      J. Biol. Chem., 2006-08-21;281(41):31164-72.
      Species: Human
      Sample Types: Whole Cells
      Applications: Bioassay
    28. Selective roles for tumor necrosis factor alpha-converting enzyme/ADAM17 in the shedding of the epidermal growth factor receptor ligand family: the juxtamembrane stalk determines cleavage efficiency.
      Authors: Hinkle CL, Sunnarborg SW, Loiselle D, Parker CE, Stevenson M, Russell WE, Lee DC
      J. Biol. Chem., 2004-04-05;279(23):24179-88.
      Species: Mouse
      Sample Types: Recombinant Protein
      Applications: Enzyme Assay
    29. Depletion of cellular cholesterol and lipid rafts increases shedding of CD30.
      Authors: von Tresckow B, Kallen KJ, von Strandmann EP, Borchmann P, Lange H, Engert A, Hansen HP
      J. Immunol., 2004-04-01;172(7):4324-31.
      Applications: Western Blot

    FAQs

    1. If the Recombinant Human TACE/ADAM17 Protein, CF (Catalog # 930-ADB) is supplied in solution consisting of 25 mM Tris, 200 mM NaCl and 0.02%(w/v) Brij-35, does the 200 mM NaCl in the formulation inhibit the activity of the protein?

      • We would not expect the 200 mM NaCl salt to affect the stability of the 930-ADB enzyme. The reconstituted material will be with DI water and is generally further diluted >1000-fold for assays, so the inhibition by NaCl becomes negligible and no impact in the activity has been observed. Ample dilution of the enzyme is necessary to ensure there are no negative effects from the salt in the formulation.

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    Recombinant Human TACE/ADAM17 Protein, CF
    By Anonymous on 10/09/2019
    Application: Enzymatic activity in vitro