Recombinant Human ADAM9 His-tag Protein, CF Summary
- R&D Systems HEK293-derived Recombinant Human ADAM9 His-tag Protein (939-ADB)
- 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
Product Specifications
Ala206-Asp697, with a C-terminal 10-His tag
Analysis
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. 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.
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 could interfere.
939-ADB
| Formulation | Supplied as a 0.2 μm filtered solution in MES 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.
|
Assay Procedure
- Assay Buffer: 25 mM Tris, 2.5 µM ZnCl2, 0.005% (w/v) Brij-35, pH 9.0
- Recombinant Human ADAM9 His-tag (rhADAM9) (Catalog # 939-ADB)
- Fluorogenic Peptide Substrate III: MCA-Pro-Leu-Ala-Gln-Ala-Val-DPA-Arg-Ser-Ser-Ser-Arg-NH3 (Catalog # ES003)
- 96-Well Black Plate
- Plate Reader with Fluorescence Read Capability
- Dilute rhADAM9 to 40 µg/mL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Combine equal volumes of 40 µg/mL rhADAM9 and 200 µM Substrate. Include a Substrate Blank by combining equal volumes of Assay Buffer and 20 µM Substrate.
- Incubate the Reactions and Substrate Blank at 37 °C for 30 minutes.
- Load 100 µL of each incubated Reaction and Substrate Blank into wells of a black plate.
- 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) |
- rhADAM9: 2 µg
- Substrate: 100 µM
Scientific Data
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2 μg/lane of Recombinant Human ADAM9 His-tag Protein (Catalog # 939-ADB) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 61-68 kDa, under reducing conditions.
Reconstitution Calculator
Background: ADAM9
Recombinant human ADAM9, also known as MDC9 or meltrin gamma, is a member of the ADAM family that contains "a disintegrin and metalloprotease like" domain (1,2). ADAM9 is widely expressed on the cell surface across multiple cell types and tissues where it is involved in diverse biological functions in the extracellular matrix (ECM) (2). Like other membrane-anchored ADAMs, expressed ADAM9 has a C-shaped structure that facilitates substrate recognition and cleavage and consists of a prodomain with a cysteine switch and furin cleavage sequence, a catalytic domain with the zinc binding site, along with several additional domains: a disintegrin-like domain, a cysteine-rich domain, an EGF-like domain, a transmembrane domain, and the cytoplasmic domain (2). A zinc ion in the catalytic domain of ADAM-9 coordinates with the prodomain to maintain latency. Removal of the propeptide through cleavage by proprotein convertases activates the protein (2). ADAM9 has specificity for targets containing an established preferred consensus sequence including a broad range of substrates within the ECM such as tumor necrosis factor-alpha (TNF-alpha), growth factors and their receptors such as the p75 TNF receptor, and key molecules beta -amyloid protein precursor, c kit ligand-1, oxidized insulin beta -chain, and fibronectin (2-4). In addition to catalytic activity, ADAM9 functions as an adhesion molecule through binding of its disintegrin domain (2,5,6) and the cytoplasmic domain of ADAM9 interacts with Src homology 3 (SH3) containing proteins and protein kinase C (2,7,8). Given the broad expression and diverse activity of ADAM9, it plays an important role in pathological diseases including degenerative diseases such as Alzheimers, inflammatory diseases such as COPD and tumor biology in many cancers including lung, prostate, liver, breast, pancreatic making it attractive as a biomarker and therapeutic target (2, 9-15).
- Moss, M.L. et al. (2001) Drug Discov. Today 6:417.
- Chou, C.W, et al. (2020) Int. J. Mol. Sci. 21:7790.
- Roghan, M. et al. (1999) J. Biol. Chem. 274:3531.
- Schwettmann, L. and H. Tschesche (2001) Prot. Express. & Purif. 21:65.
- Nath, D. et al. (2000) J. Cell Sci. 113:2319.
- Zhou, M. et al. (2001) Biochem. Biophys. Res. Comm. 280:574.
- Izumi, Y. et al. (1998) EMBO J. 17:7260.
- Howard, L. et al. (1999) J. Biol. Chem. 274:31693.
- Cong, L. and J. Jia (2011) Neurobiol. Aging 32:54.
- Zhang, J. et al. (2014) J. Cancer Res. Clin. Oncol. 140:1441.
- Hua, Y. et al. (2018) Oncol. Lett. 15:9051.
- Liu, R. et al. (2018) Onco. Targets Ther. 11:4065.
- Wang, X. et al. (2018) Am. J. Respir. Crit. Care Med. 198:1500.
- Xing, C. et al. (2019) Artif. Cells Nanomed. Biotechnol. 47:3920.
- Oh, S. et al. (2020) Cancers 12:745.
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