ADAM12, also known as meltrin-alpha, is a member of the ADAM family with metalloprotease activity (1). It consists of a propeptide, metalloproteinase, disintegrin, cysteine-rich, and EGF-like domains, a transmembrane segment, and a cytoplasmic tail with SH3 binding motifs. Human ADAM12 exists in two alternatively spliced forms: the prototype transmembrane form and a shorter secreted form lacking the transmembrane domain and the cytoplasmic tail. The secreted form has a 34 amino acid substitution in place of the transmembrane and cytoplasmic regions. In mouse, only the transmembrane form has been observed. The propeptide, which is cleaved in the Golgi by furin-like proprotein convertases, is retained in a noncovalent complex after ADAM12 secretion (2). Thus, the pro domain may function as an inhibitor of the proteolytic activity or play another unknown function. The known physiological substrates of ADAM12 are HBEGF in the heart (3) and IGFBP-3 and ‑5 in placental serum (4). Its proteolytic activity is inhibited by the tissue inhibitor of metalloproteinase-3 (Recombinant Human TIMP‑3, (Catalog # http://www.rndsystems.com/product_results.aspx?k=973-TM">973-TM) and alpha ‑2‑macroglobulin. It also mediates cell-cell adhesion by interacting with integrins and syndecans as well as with additional unidentified molecules (4). ADAM12 may be a promising marker in prenatal diagnostics and breast cancer (5, 6). The recombinant ADAM12 contains the pro, metalloproteinase, and disintegrin domains. In addition to TIMP-3, the activity can also be inhibited by 5 mM 1,10-phenanthroline.
Recombinant Human ADAM12 Protein, CF
R&D Systems | Catalog # 4416-AD
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Key Product Details
- R&D Systems CHO-derived Recombinant Human ADAM12 Protein (4416-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
CHO
Accession Number
Structure / Form
Pro & Mature forms
Applications
Enzyme Activity
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Product Specifications
Source
Chinese Hamster Ovary cell line, CHO-derived human ADAM12 protein
Arg29-Ser513, with an N-terminal signal peptide and a C-terminal 6-His tag
Arg29-Ser513, with an N-terminal signal peptide and a C-terminal 6-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
Arg29
Predicted Molecular Mass
55 kDa (Pro) & 34 kDa (Mature)
SDS-PAGE
25-30 kDa, 50-55 kDa & 68-72 kDa, reducing conditions
Activity
Measured by its ability to cleave IGFBP-3.
Reviewed Applications
Read 2 reviews rated 4 using 4416-AD in the following applications:
Formulation, Preparation, and Storage
4416-AD
| Formulation | Supplied as a 0.2 μm filtered solution in Sodium Acetate, CaCl2 and NaCl. |
| 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: ADAM12
References
- Wewer, U.M. (2004) in Handbook of Proteolytic Enzymes, Barrett, A. J. et al. eds. pp 724.
- Wewer, U.M. et al. (2006) J. Biol. Chem. 281:9481.
- Asakura, M. et al. (2002) Nat. Med. 8:35.
- Loechel, F. et al. (2000) Biochem. Biophys. Res. Comm. 278:511.
- Laigaard, J. et al. (2006) Prenat Diagn. 26:973.
- Roopali, R. et al. (2004) J. Biol. Chem. 279:51323.
Long Name
A Disintegrin and Metalloprotease-like Domain 12
Alternate Names
Meltrin alpha
Entrez Gene IDs
8038 (Human)
Gene Symbol
ADAM12
UniProt
Additional ADAM12 Products
Product Documents for Recombinant Human ADAM12 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 ADAM12 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human ADAM12 Protein, CF
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Protocols
View specific protocols for Recombinant Human ADAM12 Protein, CF (4416-AD):
Materials
- Activation Buffer: 50 mM Tris, 1 mM CaCl2, 0.5% Brij-35, pH 9.0
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, pH 7.5
- Recombinant Human ADAM12 (rhADAM12) (Catalog # 4416-AD)
- Recombinant Human Furin His-tag (rhFurin) (Catalog # 11338-SE)
- Recombinant Human IGFBP‑3 (rhIGFBP‑3) (Catalog # 675-B3)
- SDS-PAGE and/or Western blot
- Dilute rhADAM12 to 200 µg/mL in Activation Buffer.
- Dilute rhFurin to 10 µg/mL in Activation Buffer.
- Mix 25 µL of rhADAM12 and 25 µL of rhFurin in a reaction tube.
- Incubate the mixture for 1 hour at 37 °C.
- Reconstitute or dilute rhIGFBP-3 to 200 µg/mL in Assay Buffer (do not follow reconstitution directions given by the 675-B3 product insert).
- Add 25 µL of rhIGFBP-3 dilution, 10 µL of activated rhADAM12, and 15 µL of Assay Buffer to a tube.
- Incubate reaction at 37 °C for 16 hours.
- As controls combine 25 µL of rhIGFBP-3 solution and 25 µL of Assay Buffer in each of two tubes. Incubate one tube at 37 °C and the other at -20 °C.
- Stop the reaction by adding SDS-PAGE sample buffer.
- Analyze the cleavage by SDS-PAGE followed by protein staining and/or Western blot.
Per Reaction:
- rhADAM12: 20 µg/mL
- rhIGFBP-3: 100 µg/mL
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