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. Carbonic Anhydrase VB encoded by the CA5B gene is a mitochondrial protein. In comparison with another mitochondrial CA (CA5A), CA5B has different tissue distribution and chromosomal location (2, 3). The amino acid sequence of human CA5B (residues 34 to 317) is 94%, 93%, 92%, and 74% identical to that of canine, mouse, bovine/rat, and chicken.
Recombinant Human Carbonic Anhydrase VB Protein, CF
R&D Systems | Catalog # 3176-CA
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Key Product Details
- R&D Systems E. coli-derived Recombinant Human Carbonic Anhydrase VB Protein (3176-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 VB/CA5B protein
Cys34-Pro317, with a C-terminal 10-His tag
Cys34-Pro317, 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
Cys34 & Ser35
Predicted Molecular Mass
34 kDa
SDS-PAGE
34 kDa, reducing conditions
Activity
Measured by its esterase activity.
The specific activity is >150 pmol/min/µg, as measured under the described conditions.
The specific activity is >150 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
3176-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 VB/CA5B
References
- Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
- Fujikawa-Adachi, K. et al. (1999) J. Biol. Chem. 274:21228.
- Shah, G.N. et al. (2000) Proc. Natl. Acad. Sci. USA 97:1677.
Alternate Names
CA5B, CAVB
Gene Symbol
CA5B
UniProt
Additional Carbonic Anhydrase VB/CA5B Products
Product Documents for Recombinant Human Carbonic Anhydrase VB Protein, CF
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Carbonic Anhydrase VB Protein, CF
For research use only
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Protocols
View specific protocols for Recombinant Human Carbonic Anhydrase VB Protein, CF (3176-CA):
Materials
- Assay Buffer: 12.5 mM Tris, 75 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5
- Recombinant Human Carbonic Anhydrase VB (rhCA5B) (Catalog # 3176-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 rhCA5B to 2 ng/µL in Assay Buffer.
- Dilute substrate to 2 mM in Assay Buffer.
- Load in plate, 50 µL of 2 ng/µL rhCA5B, 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).
- rhCA5B: 0.1 µg
- Substrate: 1 mM
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