Recombinant Human ADAM10 is prone to proteolytic cleavage at C-terminus. The predominant form of the purified protein lacks the His tag.
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
<1.0 EU per 1 μg of the protein by the LAL method.
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.
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
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
ADAM metallopeptidase domain 10
disintegrin and metalloproteinase domain-containing protein 10
Kuzbanian protein homolog
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.
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