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.
Recombinant Human ADAM8 Protein, CF
R&D Systems | Catalog # 1031-AD
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Key Product Details
- R&D Systems NS0-derived Recombinant Human ADAM8 Protein (1031-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
NS0
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human ADAM8 protein
Met1-Pro497, with a C-terminal 6-His tag
Met1-Pro497, with a C-terminal 6-His tag
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Glu158
Predicted Molecular Mass
38 kDa
SDS-PAGE
42 kDa, reducing conditions
Activity
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.
The specific activity is >1 pmol/min/µg, as measured under the described conditions.
Reviewed Applications
Read 2 reviews rated 3.5 using 1031-AD in the following applications:
Formulation, Preparation, and Storage
1031-AD
| Formulation | Supplied as a 0.2 μm filtered solution in Glycerol, Tris, NaCl and CaCl2. |
| Shipping | 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.
|
Background: ADAM8
References
- 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.
Long Name
A Disintegrin and Metalloprotease-like Domain 8
Alternate Names
CD156a, MS2
Entrez Gene IDs
101 (Human)
Gene Symbol
ADAM8
UniProt
Additional ADAM8 Products
Product Documents for Recombinant Human ADAM8 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 ADAM8 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human ADAM8 Protein, CF
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Customer Reviews for Recombinant Human ADAM8 Protein, CF (2)
3.5 out of 5
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Application: Enzymatic activity in vitroVerified Customer | Posted 04/17/2020
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Protocols
View specific protocols for Recombinant Human ADAM8 Protein, CF (1031-AD):
Materials
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, pH 7.5 (TCN)
- Recombinant Human ADAM8 (rhADAM8) (Catalog # 1031-AD)
- Bacterial Thermolysin (Catalog # 3097-ZN)
- Phosphoramidon (Catalog # EI006), 20 mM in methanol
- Fluorogenic Peptide Substrate III: MCA-Pro-Leu-Ala-Gln-Ala-Val-DPA-Arg-Ser-Ser-Ser-Arg-NH2 (Catalog # ES003)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- 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).
Per Well:
- rhADAM8: 2.0 μg
- Substrate: 20 μM
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