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Recombinant Human ADAM9 Protein, CF Summary
Details of Functionality
Measured by its ability to cleave a fluorogenic peptide substrate Mca-PLAQAV-Dpa-RSSSR-NH2 (Catalog # ES003). The specific activity is >1.0 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human ADAM9 protein Ala206-Asp697, with a C-terminal 10-His tag
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.
Applications/Dilutions
Dilutions
Enzyme Activity
Theoretical MW
55 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
64 kDa, reducing conditions
Publications
Read Publications using 939-AD in the following applications:
Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
Dilute rhADAM9 to 40 µg/mL in Assay Buffer.
Dilute substrate to 20 µM in Assay Buffer.
Combine equal volumes of 40 µg/mL rhADAM9 and 20 µM substrate. Include a control containing Assay Buffer and 20 µM substrate without any rhADAM9.
Incubate reactions at 37 °C for 30 minutes.
Load 100 μL of reactions and control per well in a black plate.
Read with 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 Control **Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).
Per Well:
rhADAM9: 2.0 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 ADAM9 Protein, CF
a disintegrin and metalloproteinase domain 9 (meltrin gamma)
ADAM 9
ADAM metallopeptidase domain 9 (meltrin gamma)
ADAM metallopeptidase domain 9
ADAM9
Cellular disintegrin-related protein
cone rod dystrophy 9
CORD9
disintegrin and metalloproteinase domain-containing protein 9
EC 3.4.24
EC 3.4.24.-
MCMP
MCMPMDC9KIAA0021Mltng
MDC9
Meltrin gamma
Meltrin-gamma
Metalloprotease/disintegrin/cysteine-rich protein 9
MLTNG
Myeloma cell metalloproteinase
Background
ADAM9, also known as MDC9 or meltrin gamma , is a member of the ADAM family that contains a disintegrin and metalloprotease‑like domain (1). Like other membrane-anchored ADAMs, ADAM9 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. ADAM9 is able to cleave peptides corresponding to cleavage sites of tumor necrosis factor-alpha (TNF-alpha ), the p75 TNF receptor, the beta -amyloid protein precursor, and the c‑kit ligand-1, implying that it may participate in shedding of these membrane proteins (2). In fact, ADAM9 has been shown to shed membrane‑anchored heparin-binding EGF-like growth factor (3). In addition, it also cleaves oxidized insulin beta -chain and fibronectin (2,4). Besides its catalytic activity, ADAM9 functions as an adhesion molecule through binding of its disintegrin domain to integrins such as alpha v beta 5 and alpha 6 beta 1 (5, 6). The cytoplasmic domain of ADAM9 interacts with Src homology 3 (SH3)‑containing proteins and protein kinase C, and may mediate different signaling pathways (3, 7). ADAM9 is widely expressed in tissues (8).
Moss, et al. (2001) DDT 6:417.
Roghan, et al. (1999) J. Biol. Chem. 274:3531.
Izumi, et al. (1998) EMBO J. 17:7260.
Schwettmann and Tschesche (2001) Prot. Expre. & Purif. 21:65.
Nath, et al. (2000) J. Cell Sci. 113:2319.
Zhou, et al. (2001) Biochem. Biophys. Res. Comm. 280:574.
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