Carbonic Anhydrase 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. Carbonic Anhydrase VA encoded by the CA5A gene is a mitochondrial protein (2, 3). In comparison with another mitochondrial CA (CA5B), CA5A has different tissue distribution and chromosomal location (4, 5). Expression and inhibitor studies of different CAs in the rat pancreatic beta cells indicate that CA5A may be involved in the regulation of insulin secretion (6). CA5A may also participate in the detoxification of ammonia produced in the gastrointestinal tract by providing bicarbonate to carbamyl phosphate synthetase I (7). The amino acid sequence of recombinant human CA5A (residues 40 to 305) is 79%, 77%, and 76% identical to that of canine, bovine, and rat/mouse.
Recombinant Human Carbonic Anhydrase VA Protein, CF
R&D Systems | Catalog # 3049-CA
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Key Product Details
- R&D Systems E. coli-derived Recombinant Human Carbonic Anhydrase VA Protein (3049-CA)
- 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
E. coli
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
E. coli-derived human Carbonic Anhydrase VA/CA5A protein
Ala40-Ser305, with a C-terminal 10-His tag
Ala40-Ser305, with a C-terminal 10-His tag
Purity
>95%, 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
Ala40
Predicted Molecular Mass
31 kDa
SDS-PAGE
32 kDa, reducing conditions
Activity
Measured by its esterase activity.
The specific activity is >100 pmol/min/µg, as measured under the described conditions.
The specific activity is >100 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
3049-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.
|
Background: Carbonic Anhydrase VA/CA5A
References
- Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
- Nagao, Y. et al. (1993) Proc. Natl. Acad. Sci. USA 90:7623.
- Nagao, Y. et al. (1995) Genomics 28:477.
- Shah, G.N. et al. (2000) Proc. Natl. Acad. Sci. USA 97:1677.
- Fujikawa-Adachi, K. et al. (1999) J. Biol. Chem. 274:21228.
- Parkkila, A.K. et al. (1998) J. Biol. Chem. 273:24620.
- Saarnio, J. et al. (1999) J. Histochem. Cytochem. 47:517.
Alternate Names
CA5A, CAVA
Gene Symbol
CA5A
UniProt
Additional Carbonic Anhydrase VA/CA5A Products
Product Documents for Recombinant Human Carbonic Anhydrase VA Protein, CF
Product Specific Notices for Recombinant Human Carbonic Anhydrase VA Protein, CF
For research use only
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Protocols
View specific protocols for Recombinant Human Carbonic Anhydrase VA Protein, CF (3049-CA):
Materials
- Assay Buffer: 12.5 mM Tris, 75 mM NaCl, 0.05% (v/v) Brij-35, pH 7.5
- Recombinant Human Carbonic Anhydrase VA (rhCA5A) (Catalog # 3049-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 rhCA5A 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 rhCA5A, and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing 50 µL of 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).
- rhCA5A: 1 µg
- Substrate: 1 mM