Recombinant Human Carbonic Anhydrase IX Protein, CF Summary
Pro59-Asp414, with a C-terminal 10-His tag
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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.
|Formulation||Lyophilized from a 0.2 μm filtered solution in Tris and NaCl.|
|Reconstitution||Reconstitute at 100 μg/mL in sterile 25 mM Tris and 150 mM NaCl, pH 7.5.|
|Shipping||The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.|
|Stability & Storage:||Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- Assay Buffer: 12.5 mM Tris, 75 mM NaCl, pH 7.5
- Recombinant Human Carbonic Anhydrase IX/CA9 (rhCA9) (Catalog # 2188-CA)
- Substrate: 4-Nitrophenyl acetate (4-NPA) (Sigma, Catalog # N8130), 100 mM stock in acetone
- 96-well Clear Plate (Costar, Catalog # 2592)
- Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rhCA9 to 20 ng/µL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- Load in plate, 50 µL of 20 ng/µL rhCA9, and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 2 mM Substrate.
- Read at a wavelength of 400 nm (bottom read) in kinetic mode for 5 minutes.
- Calculate specific activity:
Specific Activity (pmol/min/µg) =
|Adjusted Vmax* (OD/min) x Conversion Factor** (pmol/OD)|
|amount of enzyme (µg)|
*Adjusted for Substrate Blank
**Derived using calibration standard 4-Nitrophenol (Sigma, Catalog # 241326).Per Well:
- rhCA9: 1 µg
- Substrate: 1 mM
Background: Carbonic Anhydrase IX/CA9
Carbonic Anhydrase (CA) catalyzes the reversible reaction of CO2 + H2O = HCO3- + H+, which is fundamental to many processes such as respiration, renal tubular acidification and bone resorption (1-3). Topics in the CA meeting (6th International Conference on the CAs, June 20‑25, 2003, Slovakia) ranged from use of CAs as markers for tumor and hypoxia in clinic, as nutritional supplement in milk, and as a tool for CO2 removal and mosquito control in industry. CA9, also known as membrane antigen MN and renal cell carcinoma (RCC)‑associated antigen G250, is a transmembrane enzyme expressed primarily in carcinoma cells. It is one of the best markers for hypoxia and for RCC (4, 5). rhCA9 corresponds to the extracellular portion of human CA9.
- Pastorek, J. et al. (1994) Oncogene 9:2877.
- Opavsky, R. et al. (1996) Genomics 33:480.
- Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
- Kaluzova, M. et al. (2004) Mol. Cell Biol. 24:5757.
- Mukouyama, H. et al. (2004) Clin. Cancer Res. 10:1421.
Citations for Recombinant Human Carbonic Anhydrase IX Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
Citations: Showing 1 - 4
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Design, synthesis and biological evaluation of novel carbohydrate-based sulfonamide derivatives as antitumor agents
Authors: S Hao, X Cheng, X Wang, R An, H Xu, M Guo, C Li, Y Wang, Z Hou, C Guo
Bioorg. Chem., 2020;104(0):104237.
Sample Types: Peptide
The interactome of metabolic enzyme carbonic anhydrase IX reveals novel roles in tumor cell migration and invadopodia/MMP14-mediated invasion
Authors: M Swayampaku, PC McDonald, M Vallejo, E Coyaud, SC Chafe, A Westerback, G Venkateswa, J Shankar, G Gao, EMN Laurent, Y Lou, KL Bennewith, CT Supuran, IR Nabi, B Raught, S Dedhar
Slowing down glioblastoma progression in mice by running or the anti-malarial drug dihydroartemisinin? Induction of oxidative stress in murine glioblastoma therapy
Authors: D Lemke, HW Pledl, M Zorn, M Jugold, E Green, J Blaes, S Löw, A Hertenstei, M Ott, F Sahm, AC Steffen, M Weiler, F Winkler, M Platten, Z Dong, W Wick
Measuring carbonic anhydrase IX as a hypoxia biomarker: differences in concentrations in serum and plasma using a commercial enzyme-linked immunosorbent assay due to influences of metal ions.
Authors: Wind TC, Messenger MP, Thompson D, Selby PJ, Banks RE
Ann. Clin. Biochem., 2011;48(0):112-20.
Sample Types: N/A
Applications: ELISA (Control)
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