Recombinant Human Carbonic Anhydrase XII Protein, CF
Recombinant Human Carbonic Anhydrase XII Protein, CF Summary
Product Specifications
Ala25-Gln291, 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.
2190-CA
| Formulation | Supplied as a 0.2 μm filtered solution in Tris 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: 12.5 mM Tris, 75 mM NaCl, pH 7.5
- Recombinant Human Carbonic Anhydrase XII/CA12 (rhCA12) (Catalog # 2190-CA)
- Substrate: 4-Nitrophenyl Acetate (4-NPA) (Sigma, Catalog # N8130), 100 mM stock in acetone
- 96-well Clear Plate (Costar, Catalog # 92592)
- Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rhCA12 to 20 ng/µL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- In a plate, load 50 µL of 20 ng/µL rhCA12, and start the reaction by adding 50 µL of 2 mM Substrate to wells. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 2 mM Substrate.
- Read absorbance 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).
- rhCA12: 1 µg
- Substrate: 1 mM
Reconstitution Calculator
Background: Carbonic Anhydrase XII/CA12
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). Topics in a CA meeting (6th International Conference on the CAs, June 20‑25, 2003, Slovakia) ranged from the use of CAs as markers for tumor and hypoxia in the clinic, as a nutritional supplement in milk, and as a tool for CO2 removal and mosquito control in industry. CA12 is a type I membrane enzyme highly expressed in colon, kidney, prostate, intestine and activated lymphocytes and moderately expressed in pancreas, ovary, and testis (2, 3). It is over-expressed in some renal cell cancers and in glaucoma (2, 4). Two alternatively spliced forms exist, which either contains or lacks a 11 amino acid segment just before the transmembrane domain (5). The secreted, purified recombinant human CA12 contains the residues that are common for both forms.
- Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
- Tuereci, O. et al. (1998) Proc. Natl. Acad. Sci. USA 95:7608.
- Ivanov, S.V. et al. (1998) Proc. Natl. Acad. Sci. USA 95:12596.
- Liao, S.-Y. et al. (2003) J. Med. Genet. 40:257.
- Strausberg, R.L. et al. (2002) Proc. Natl. Acad. Sci. USA 99:16899.
Citation for Recombinant Human Carbonic Anhydrase XII 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.
1 Citation: Showing 1 - 1
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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., 2020-08-27;104(0):104237.
Species: Human
Sample Types: Peptide
Applications: Bioassay
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