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Recombinant Human ADAM10 Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

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Recombinant Human ADAM10 Protein, CF Summary

Details of Functionality
Measured by its ability to cleave a fluorogenic peptide substrate Mca-KPLGL-Dpa-AR-NH2 (Catalog # ES010). The specific activity is >20 pmol/min/µg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human ADAM10 protein
Thr214-Glu672, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Thr214
Structure / Form
Recombinant Human ADAM10 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
ADAM10
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
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
60 kDa, reducing conditions
Publications
Read Publications using
936-AD 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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in MES, NaCl, ZnCl2,Glycerol and Brij-35.
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
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
  1. Dilute rhADAM10 to 1 ng/µL in Assay Buffer.
  2. Dilute substrate to 20 µM in Assay Buffer.
  3. 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.
  4. Seal plate and incubate at 37 °C for 30 minutes.
  5. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in endpoint mode.
  6. 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).

Per Well:
  • rhADAM10: 0.050 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 ADAM10 Protein, CF

  • a disintegrin and metalloprotease domain 10
  • a disintegrin and metalloproteinase domain 10
  • AD10
  • ADAM 10
  • ADAM metallopeptidase domain 10
  • ADAM10
  • CD156c antigen
  • CD156c
  • CDw156
  • disintegrin and metalloproteinase domain-containing protein 10
  • EC 3.4.24
  • HsT18717
  • kuz
  • Kuzbanian protein homolog
  • Kuzbanian
  • MADM
  • MADMEC 3.4.24.81
  • Mammalian disintegrin-metalloprotease

Background

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).

  1. Rooke, et al. (1996) Science 273:1227.
  2. Pan and Rubin (1997) Cell 90:271.
  3. Qi, et al. (1999) Science 283:91.
  4. Lammich, et al. (1999) Proc. Natl. Acad. Sci. USA 96:3922.
  5. Rosendahl, et al. (1997) J. Biol. Chem. 272:24588.
  6. Chantry, et al. (1989) J. Biol. Chem. 264:21603.
  7. Millichip, et al. (1998) Biochem. Biophys. Res. Comm. 245:594.
  8. Chantry and Glynn (1990) Biochem. J. 268:245.
  9. Fahrenholz, et al. (2000) Ann. N.Y. Acad. Sci. 920:215.

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Applications: Enzyme Activity

Publications for ADAM10 (936-AD)(24)

We have publications tested in 4 confirmed species: Human, Mouse, Bacteria - E. Coli, Hamster.

We have publications tested in 5 applications: Bioassay, Enzyme Assay, In Vivo, Inhibition, Surface Plasmon Resonance.


Filter By Application
Bioassay
(14)
Enzyme Assay
(7)
In Vivo
(1)
Inhibition
(1)
Surface Plasmon Resonance
(1)
All Applications
Filter By Species
Human
(17)
Mouse
(3)
Bacteria - E. Coli
(1)
Hamster
(1)
All Species
Showing Publications 1 - 10 of 24. Show All 24 Publications.
Publications using 936-AD Applications Species
J Segura, J Ireland, Z Zou, G Roth, J Buchwald, TJ Shen, E Fischer, S Moir, TW Chun, PD Sun HIV-1 release requires Nef-induced caspase activation PLoS ONE, 2023-02-13;18(2):e0281087. 2023-02-13 [PMID: 36780482] (Bioassay, Human) Bioassay Human
M Zatovicova, I Kajanova, M Takacova, L Jelenska, O Sedlakova, M Labudova, S Pastorekov ADAM10 mediates shedding of carbonic anhydrase IX ectodomain non?redundantly to ADAM17 Oncology reports, 2022-12-16;49(2):. 2022-12-16 [PMID: 36524367] (Bioassay, Human) Bioassay Human
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 A topological refactoring design strategy yields highly stable granulopoietic proteins Nature Communications, 2022-05-26;13(1):2948. 2022-05-26 [PMID: 35618709] (Bioassay, Human) Bioassay Human
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 Altered succinylation of mitochondrial proteins, APP and tau in Alzheimer&#039;s disease Nature Communications, 2022-01-10;13(1):159. 2022-01-10 [PMID: 35013160] (Bioassay, Human) Bioassay Human
Q Renfeng, C Shuxiao, G Peixian, L Kun, F Xuedong, Y Hai, W Xuejun, L Gang ADAM10 attenuates the development of abdominal aortic aneurysms in a mouse model Molecular Medicine Reports, 2021-09-07;24(5):. 2021-09-07 [PMID: 34490486] (In Vivo, Mouse) In Vivo Mouse
AP Carreca, VM Pravatà, M Markham, S Bonelli, G Murphy, H Nagase, L Troeberg, SD Scilabra TIMP-3 facilitates binding of target metalloproteinases to the endocytic receptor LRP-1 and promotes scavenging of MMP-1 Sci Rep, 2020-07-21;10(1):12067. 2020-07-21 [PMID: 32694578] (Surface Plasmon Resonance, Human) Surface Plasmon Resonance 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., 2019-10-18;0(0):103365. 2019-10-18 [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, 2019-10-01;9(1):14086. 2019-10-01 [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., 2019-05-06;130(0):. 2019-05-06 [PMID: 31063986] (Bioassay, Human) Bioassay 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, 2018-06-23;9(14):2559-2570. 2018-06-23 [PMID: 30026855] (Bioassay) Bioassay
Show All 24 Publications.

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Bioinformatics

Gene Symbol ADAM10
Uniprot