Recombinant Human Carbonic Anhydrase XIV Protein, CF

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

Recombinant Human 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 human Carbonic Anhydrase XIV/CA14 protein
  Gly19-Met290, with a C-terminal 10-His tag
• Accession #: Q9ULX7
• N-terminal Sequence: Gly19
• Protein/Peptide Type: Recombinant Enzymes
• Gene: CA14
• 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 Human Carbonic Anhydrase XIV/CA14 (rhCA14) (Catalog # 2195-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
  1. Dilute rhCA14 to 8 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. In a plate, load 50 µL of 8 ng/µL rhCA14, and start the reaction by adding 50 µL of 2 mM Substrate to wells.
  4. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 2 mM Substrate.
  5. Read absorbance at wavelength of 400 nm (bottom read) in kinetic mode for 5 minutes.
  6. 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:
  • rhCA14: 0.4 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 Human 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 the use of CAs as markers for tumor and hypoxia in the clinic, as anutritional supplement in milk, and as a tool for CO₂ removal and mosquito control in industry. CA14 is a type I membrane enzyme highly expressed in all parts of the central nervous system with lower expression in adult liver, heart, small intestine, colon, kidney, urinary bladder, and skeletal muscle (2).

1. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
2. Fujikawa-Adachi, K. et al. (1999) Genomics 61:74.

Bioinformatics

• Gene Symbol: CA14
• Uniprot:
  • Q9ULX7()