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).
Recombinant Human ADAM10 Protein, CF
R&D Systems | Catalog # 936-AD
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Key Product Details
- R&D Systems Sf 21 (baculovirus)-derived Recombinant Human ADAM10 Protein (936-AD)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
Sf 21 (baculovirus)
Accession Number
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.
Applications
Enzyme Activity
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Product Specifications
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human ADAM10 protein
Thr214-Glu672, with a C-terminal 10-His tag
Thr214-Glu672, with a C-terminal 10-His tag
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Thr214
Predicted Molecular Mass
52 kDa
SDS-PAGE
60 kDa, reducing conditions
Activity
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.
The specific activity is >20 pmol/min/µg, as measured under the described conditions.
Reviewed Applications
Read 1 review rated 5 using 936-AD in the following applications:
Formulation, Preparation, and Storage
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.
|
Background: ADAM10
References
- 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.
Long Name
A Disintegrin and Metalloprotease-like Domain 10
Alternate Names
CD156c, Kuzbanian, MADM
Gene Symbol
ADAM10
UniProt
Additional ADAM10 Products
Product Documents for Recombinant Human ADAM10 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human ADAM10 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human ADAM10 Protein, CF
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Protocols
View specific protocols for Recombinant Human ADAM10 Protein, CF (936-AD):
Materials
- 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).
Per Well:
- rhADAM10: 0.050 µg
- Substrate: 10 µM
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Associated Pathways
