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 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. Mouse CA14 is a type I membrane enzyme highly expressed in the kidney and heart, followed by the skeletal muscle, brain, lung and liver (2). The deduced amino acid sequence of mouse CA14 consists of a signal peptide (residues 1 to 15), an ectodomain (residues 16 to 290), a transmembrane domain (residues 291 to 311) and a cytoplamic region (residues 32 to 337). rmCA14 corresponds to the ectodomain and has activity as described in Activity Assay Protocol.
Recombinant Mouse Carbonic Anhydrase XIV Protein, CF
R&D Systems | Catalog # 2504-CA
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Key Product Details
- R&D Systems NS0-derived Recombinant Mouse Carbonic Anhydrase XIV Protein (2504-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
NS0
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived mouse Carbonic Anhydrase XIV/CA14 protein
Ala16-Met290, with a C-terminal 10-His tag
Ala16-Met290, 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
Ala16
Predicted Molecular Mass
32 kDa
SDS-PAGE
43-54 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
2504-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 XIV/CA14
References
- Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
- Mori, K. et al. (1999) J. Biol. Chem. 274:15701.
Alternate Names
CA14
Gene Symbol
CA14
UniProt
Additional Carbonic Anhydrase XIV/CA14 Products
Product Documents for Recombinant Mouse Carbonic Anhydrase XIV Protein, CF
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Mouse Carbonic Anhydrase XIV Protein, CF
For research use only
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Protocols
View specific protocols for Recombinant Mouse Carbonic Anhydrase XIV Protein, CF (2504-CA):
Materials
- Assay Buffer: 12.5 mM Tris, 75 mM NaCl, pH 7.5
- Recombinant Mouse Carbonic Anhydrase XIV/CA14 (rmCA14) (Catalog # 2504-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: Spectra-Max Plus by Molecular Devices) or equivalent
- Dilute rmCA14 to 16 ng/µL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- Load in a plate 50 µL of 16 ng/µL rmCA14, 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).
- rmCA14: 0.8 µg
- Substrate: 1 mM
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