Recombinant Human ADAM10 Protein, CF Summary
Product Specifications
Thr214-Glu672, with a C-terminal 10-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
936-AD
Formulation | Supplied as a 0.2 μm filtered solution in MES, NaCl, ZnCl2,Glycerol and Brij-35. |
Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Assay Procedure
- Assay Buffer: 25 mM Tris, 2 μM ZnCl2, 0.005% (w/v) Brij-35, pH 9.0
- Recombinant Human ADAM10 (rhADAM10) (Catalog # 936-AD)
- Fluorogenic Peptide Substrate: MCA-Lys-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # ES010), 6.2 mM in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhADAM10 to 1 ng/µL in Assay Buffer.
- Dilute substrate to 20 µM in Assay Buffer.
- Load into plate 50 µL of 1 ng/µL rhADAM10 and start the reaction by adding 50 µL of 20 µM Substrate. As a control load 50 µL of Assay Buffer and 50 µL of 20 µM Substrate.
- Seal plate and incubate at 37 °C for 30 minutes.
- Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in endpoint mode.
- Calculate specific activity:
Specific Activity (pmol/min/µg) = |
Adjusted Fluorescence* (RFU) x Conversion Factor** (pmol/RFU) |
Incubation time (min) x amount of enzyme (µg) |
*Adjusted for Substrate Blank.
**Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).
- rhADAM10: 0.050 µg
- Substrate: 10 µM
Reconstitution Calculator
Background: ADAM10
ADAM10 (also known as Kuzbanian, mammalian disintegrin metalloprotease, myelin-associated metalloproteinase) is a member of the ADAM family that contains a disintegrin and metalloprotease-like domain (1, 2). Like other membrane-anchored ADAMs, ADAM10 consists of the following domains, pro with a cysteine switch and furin cleavage sequence, catalytic with the zinc-binding site and Met-turn expected for reprolysins, disintegrin-like, cysteine-rich, EGF-like, transmembrane, and cytoplasmic. ADAM10 is highly conserved, with 97% amino acid identity between mouse, rat, bovine and human and 45% identity between mouse and Drosophila. The active enzyme processes notch, notch ligand delta, and amyloid protein precursor at the alpha site, playing an important role in neurogenesis (3, 4). It also processes the 26 kDa membrane-anchored pro-tumor necrosis factor-alpha (TNF-alpha ) to the 17 kDa mature TNF-alpha (5). It cleaves myelin basic protein and type IV collagen (6, 7). ADAM10 is widely expressed in tissues and resides both on the cell surface and in the cell (8, 9).
- Rooke, et al. (1996) Science 273:1227.
- Pan and Rubin (1997) Cell 90:271.
- Qi, et al. (1999) Science 283:91.
- Lammich, et al. (1999) Proc. Natl. Acad. Sci. USA 96:3922.
- Rosendahl, et al. (1997) J. Biol. Chem. 272:24588.
- Chantry, et al. (1989) J. Biol. Chem. 264:21603.
- Millichip, et al. (1998) Biochem. Biophys. Res. Comm. 245:594.
- Chantry and Glynn (1990) Biochem. J. 268:245.
- Fahrenholz, et al. (2000) Ann. N.Y. Acad. Sci. 920:215.
Citations for Recombinant Human ADAM10 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.
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Citations: Showing 1 - 10
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HIV-1 release requires Nef-induced caspase activation
Authors: J Segura, J Ireland, Z Zou, G Roth, J Buchwald, TJ Shen, E Fischer, S Moir, TW Chun, PD Sun
PLoS ONE, 2023-02-13;18(2):e0281087.
Species: Human
Sample Types: Peptide
Applications: Bioassay -
ADAM10 mediates shedding of carbonic anhydrase IX ectodomain non?redundantly to ADAM17
Authors: M Zatovicova, I Kajanova, M Takacova, L Jelenska, O Sedlakova, M Labudova, S Pastorekov
Oncology reports, 2022-12-16;49(2):.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
A topological refactoring design strategy yields highly stable granulopoietic proteins
Authors: J Skokowa, B Hernandez, M Coles, M Ritter, M Nasri, J Haaf, N Aghaallaei, Y Xu, P Mir, AC Krahl, KW Rogers, K Maksymenko, B Bajoghli, K Welte, AN Lupas, P Müller, M ElGamacy
Nature Communications, 2022-05-26;13(1):2948.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
Altered succinylation of mitochondrial proteins, APP and tau in Alzheimer's disease
Authors: Y Yang, V Tapias, D Acosta, H Xu, H Chen, R Bhawal, ET Anderson, E Ivanova, H Lin, BT Sagdullaev, J Chen, WL Klein, KL Viola, S Gandy, V Haroutunia, MF Beal, D Eliezer, S Zhang, GE Gibson
Nature Communications, 2022-01-10;13(1):159.
Species: Human
Sample Types: Peptides
Applications: Bioassay -
ADAM10 attenuates the development of abdominal aortic aneurysms in a mouse model
Authors: Q Renfeng, C Shuxiao, G Peixian, L Kun, F Xuedong, Y Hai, W Xuejun, L Gang
Molecular Medicine Reports, 2021-09-07;24(5):.
Species: Mouse
Sample Types: In Vivo
Applications: In Vivo -
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 -
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 -
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 -
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 -
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 -
Novel therapeutic features of disulfiram against hepatocellular carcinoma cells with inhibitory effects on a disintegrin and metalloproteinase 10
Authors: K Goto, J Arai, A Stephanou, N Kato
Oncotarget, 2018-04-10;9(27):18821-18831.
Applications: Enzyme Assay -
Non-cell-autonomous function of DR6 in Schwann cell proliferation
Authors: A Colombo, HE Hsia, M Wang, PH Kuhn, MS Brill, P Canevazzi, R Feederle, C Taveggia, T Misgeld, SF Lichtentha
EMBO J., 2018-02-19;0(0):.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
Matrix metalloproteinase activity stimulates N-cadherin shedding and the soluble N-cadherin ectodomain promotes classical microglial activation
Authors: K Conant, S Daniele, PL Bozzelli, T Abdi, A Edwards, A Szklarczyk, I Olchefske, D Ottenheime, K Maguire-Ze
J Neuroinflammation, 2017-03-17;14(1):56.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
Harnessing the natural inhibitory domain to control TNF? Converting Enzyme (TACE) activity in vivo
Sci Rep, 2016-12-16;6(0):35598.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay -
Serological immune response against ADAM10 pro-domain is associated with favourable prognosis in stage III colorectal cancer patients
Authors: SM Álvarez-Fe, M Barbariga, L Cannizzaro, CV Cannistrac, L Hurley, A Zanardi, A Conti, F Sanvito, A Innocenzi, N Pecorelli, M Braga, M Alessio
Oncotarget, 2016-11-29;7(48):80059-80076.
Species: Human
Sample Types: Serum
Applications: Enzyme Assay -
Growth factor and co-receptor release by structural regulation of substrate metalloprotease accessibility
Sci Rep, 2016-11-23;6(0):37464.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay -
Stanniocalcin-1 Potently Inhibits the Proteolytic Activity of the Metalloproteinase Pregnancy-associated Plasma Protein-A.
Authors: Kloverpris S, Mikkelsen J, Pedersen J, Jepsen M, Laursen L, Petersen S, Oxvig C
J Biol Chem, 2015-07-20;290(36):21915-24.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay -
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 -
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 -
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 -
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 -
Anticancer chemotherapy inhibits MHC class I-related chain a ectodomain shedding by downregulating ADAM10 expression in hepatocellular carcinoma.
Authors: Kohga K, Takehara T, Tatsumi T, Miyagi T, Ishida H, Ohkawa K, Kanto T, Hiramatsu N, Hayashi N
Cancer Res., 2009-10-13;69(20):8050-7.
Species: Human
Sample Types: Whole Tissue
Applications: Bioassay -
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 -
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
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