Recombinant Human Carbonic Anhydrase XII Protein, CF

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

Recombinant Human Carbonic Anhydrase XII Protein, CF Summary

• Details of Functionality: Measured by its esterase activity. The specific activity is >10 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived human Carbonic Anhydrase XII/CA12 protein
  Ala25-Gln291, with a C-terminal 10-His tag
• Accession #: O43570
• N-terminal Sequence: Ala25
• Protein/Peptide Type: Recombinant Enzymes
• Gene: CA12
• 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: 38-41 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 Human Carbonic Anhydrase XII/CA12 (rhCA12) (Catalog # 2190-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 rhCA12 to 20 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. In a plate, load 50 µL of 20 ng/µL rhCA12, and start the reaction by adding 50 µL of 2 mM Substrate to wells. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 2 mM Substrate.
  4. Read absorbance 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:
  • rhCA12: 1 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 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 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. CA12 is a type I membrane enzyme highly expressed in colon, kidney, prostate, intestine and activated lymphocytes and moderately expressed in pancreas, ovary, and testis (2, 3). It is over-expressed in some renal cell cancers and in glaucoma (2, 4). Two alternatively spliced forms exist, which either contains or lacks a 11 amino acid segment just before the transmembrane domain (5). The secreted, purified recombinant human CA12 contains the residues that are common for both forms.

1. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
2. Tuereci, O. et al. (1998) Proc. Natl. Acad. Sci. USA 95:7608.
3. Ivanov, S.V. et al. (1998) Proc. Natl. Acad. Sci. USA 95:12596.
4. Liao, S.-Y. et al. (2003) J. Med. Genet. 40:257.
5. Strausberg, R.L. et al. (2002) Proc. Natl. Acad. Sci. USA 99:16899.

Publications for Carbonic Anhydrase XII/CA12 (2190-CA)(1)

Publication using 2190-CA

• S Hao, X Cheng, X Wang, R An, H Xu, M Guo, C Li, Y Wang, Z Hou, C Guo Design, synthesis and biological evaluation of novel carbohydrate-based sulfonamide derivatives as antitumor agents Bioorg. Chem., 2020-08-27;104(0):104237. 2020-08-27 [PMID: 32911194] (Bioassay, Human)

Bioinformatics

• Gene Symbol: CA12
• Uniprot:
  • O43570()