Mouse myeloma cell line, NS0-derived human ADAM8 protein Met1-Pro497, with a C-terminal 6-His tag
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis
Predicted Molecular Mass
42 kDa, reducing conditions
Measured by its ability to cleave a fluorogenic peptide substrate Mca-PLAQAV-Dpa-RSSSR-NH2 (Catalog # ES003). The specific activity is >1 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation and Storage
What does CF mean?
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our
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.
What formulation is right for me?
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
Supplied as a 0.2 μm filtered solution in Glycerol, Tris, NaCl and CaCl2.
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
6 months from date of receipt, -20 to -70 °C as supplied.
3 months, -20 to -70 °C under sterile conditions after opening.
Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, pH 7.5 (TCN)
Recombinant Human ADAM8 (rhADAM8) (Catalog # 1031-AD)
Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
Dilute rhADAM8 to 400 μg/mL in Assay Buffer.
Combine equal volumes 1.5 μg/mL thermolysin and diluted rhADAM8 so that the final concentration of rhADAM8 is 200 μg/mL and thermolysin is 0.75 μg/mL.
Incubate at 37 °C for 30 minutes.
Stop reaction by adding Phosphoramidon to a final concentration of 0.05 mM.
Incubate at room temperature for 15 minutes.
Dilute activated rhADAM8 to 40 ng/μL in Assay Buffer.
Dilute substrate to 40 μM in Assay Buffer.
Load 50 μL of 40 ng/μL rhADAM8 in a black well plate and start the reaction by adding 50 μL of 40 μM substrate. Include a control containing 50 μL Assay Buffer and 50 μL of 40 μM substrate.
Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
Calculate specific activity:
Specific Activity (pmol/min/µg) =
Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)
*Adjusted for Substrate Blank **Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).
rhADAM8: 2.0 μg
Substrate: 20 μM
ADAM8, also known as cell surface antigen MS2 or CD156a, is a member of the ADAM family that contains a disintegrin and metalloprotease-like domain (1, 2). ADAM8 can cleave a variety of substrates and has been shown as a sheddase for the low affinity IgE receptor CD23 and the neural recognition molecule CHL1 (3, 4). Expression and regulation studies suggest that ADAM8 is a novel osteoclast stimulating factor and may play a role in asthma (5, 6). The 824 amino acid precursor of human ADAM8 consists of a signal peptide (residues 1 to 16), a pro peptide (residues 17 to 199), a metaloprotease domain (residues 200 to 400), a disintegrin-like domain (residues 408 to 494), a cysteine-rich region (residues 497 to 613), an EGF-like domain (residues 614 to 640), a transmembrane region (residues 656 to 676) and a cytoplasmic domain (residues 677 to 824). The purified rhADAM8 (residues 158 to 497) contains a part of the pro peptide and the entire metalloprotease and disintegrin-like domains. It can be activated and assayed under the conditions described in the Activity Assay Protocol.
Yoshiyama, K. et al. (1997) Genomics 41:56.
Moss, M.L. and J.W. Bartsch (2004) Biochemistry 43:7227.
Fourie, A.M. et al. (2003) J. Biol. Chem. 278:30469.
Naus, S. et al. (2004) J. Biol. Chem. 279:16083.
Choi, S.J. et al. (2001) J. Bone Miner. Res. 16:814.
King, N.E. et al. (2004) Am. J. Respir. Cell Mol. Biol. 31:257.
A Disintegrin and Metalloprotease-like Domain 8
Entrez Gene IDs
ADAM 8, ADAM metallopeptidase domain 8, ADAM8, CD156a, CD156a antigen, CD156human leukocyte differentiation antigen, Cell surface antigen MS2, EC 3.4.24, EC 3.4.24.-, MGC134985, MS2, MS2disintegrin and metalloproteinase domain-containing protein 8, a disintegrin and metalloproteinase domain 8
Which Brands are Currently Available on bio-techne.com? R&D Systems, Tocris Bioscience and ProteinSimple branded products are available to purchase through bio‑techne.com. ProteinSimple branded instruments and Simple Plex Assays are available to quote. ProteinSimple branded instrument consumables and ACD branded products will be available on bio-techne.com in the near future. Novus Biologicals branded products are not currently available on bio‑techne.com and can be found at novusbio.com.
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