Recombinant Mouse Carbonic Anhydrase XIV Protein, CF

• Reactivity: Mu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Mouse Carbonic Anhydrase XIV Protein, CF Summary

• Details of Functionality: Measured by its esterase activity. The specific activity is >100 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived mouse Carbonic Anhydrase XIV/CA14 protein
  Ala16-Met290, with a C-terminal 10-His tag
  Accession # Q9WVT6
• Accession #: Q9WVT6
• N-terminal Sequence: Ala16
• Protein/Peptide Type: Recombinant Enzymes
• Gene: Car14
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 32 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 43 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
• Buffer: Supplied as a 0.2 μm filtered solution in Tris and NaCl.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Assay Procedure:
  • 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
  1. Dilute rmCA14 to 16 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. 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.
  4. Read at a wavelength of 400 nm (bottom read) in kinetic mode for 5 minutes.
  5. 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:
  • rmCA14: 0.8 µg
  • Substrate: 1 mM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse Carbonic Anhydrase XIV Protein, CF

• CA14
• Carbonate dehydratase XIV
• Carbonic Anhydrase XIV
• carbonic anhydrase XIVcarbonic anhydrase 14
• carbonic dehydratase
• CAXiV
• CA-XIV
• EC 4.2.1.1

Background

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 (6^th 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.

1. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50. 
2. Mori, K. et al. (1999) J. Biol. Chem. 274:15701.

Bioinformatics

• Gene Symbol: Car14
• Uniprot:
  • Q9WVT6()