Recombinant Mouse Carbonic Anhydrase XII Protein, CF

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

Recombinant Mouse Carbonic Anhydrase XII Protein, CF Summary

• Details of Functionality: Measured by its esterase activity. The specific activity is >50 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived mouse Carbonic Anhydrase XII/CA12 protein
  Ala25-Ser301, with a C-terminal 10-His tag
  Accession # Q8CI85
• Accession #: Q8CI85
• N-terminal Sequence: Ala25
• Protein/Peptide Type: Recombinant Enzymes
• Gene: Car12
• 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: 33 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: 39-41 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 XII/CA12 (rmCA12) (Catalog # 2345-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 rmCA12 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 rmCA12 and include a Substrate Blank containing 50 µL Assay Buffer.
  4. Start the reaction by adding 50 µL of 2 mM Substrate to wells.
  5. Read at a 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:
  • rmCA12: 0.40 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 XII Protein, CF

• CA12
• Carbonate dehydratase XII
• carbonic anhydrase 12
• Carbonic Anhydrase XII
• carbonic anhydrase XIICAXII
• carbonic dehydratase
• CA-XII
• EC 4.2.1.1
• FLJ20151
• HsT18816
• Tumor antigen HOM-RCC-3.1.3

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 CA12 is predicted to be a type I membrane enzyme (2, 3). The secreted, purified recombinant mouse CA12 corresponds to the extracellular domain and has both CA activity and esterase activity. To our knowledge, this is the first description of purified recombinant mouse CA12 with enzymatic activity.

1. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
2. Strausberg, R.L. et al. (2002) Proc. Natl. Acad. Sci. USA 99:16899.
3. Okazaki, Y. et al. (2002) Nature 420:563.

Bioinformatics

• Gene Symbol: Car12
• Uniprot:
  • Q8CI85()