Recombinant Human Carbonic Anhydrase VII Protein, CF

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

Recombinant Human Carbonic Anhydrase VII Protein, CF Summary

• Details of Functionality: Measured by its esterase activity. The specific activity is >20 pmol/min/µg, as measured under the described conditions.
• Source: E. coli-derived human Carbonic Anhydrase VII/CA7 protein
  Thr2-Ala264, with a C-terminal 10-His tag
• Accession #: NP_005173
• N-terminal Sequence: Thr2
• Protein/Peptide Type: Recombinant Enzymes
• Gene: CA7
• 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: 31 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: 31 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 VII/CA7 (rhCA7) (Catalog # 3177-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 rhCA7 to 40 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Load in plate, 50 µL of 50 ng/µL rhCA7, and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing 50 µL 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:
  • rhCA7: 2 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 VII Protein, CF

• CA7
• Carbonate dehydratase VII
• carbonic anhydrase 7
• Carbonic Anhydrase VII
• carbonic anhydrase VIICAVII
• carbonic dehydratase VII
• CA-VII
• EC 4.2.1.1

Background

Carbonic anhydrase catalyzes the reversible reaction of CO₂ + H₂O = HCO₃^- + H⁺, which is fundamental to many processes such as respiration, renal tubular acidification and bone resorption (2). 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. Carbonic Anhydrase VII encoded by the CA7 gene is a cytosolic protein predominantly expressed in the salivary gland (1). CA7 may have additional tissue distributions and functions. For example, studies with CA inhibitors provide evidence that human CA7 is the CA isozyme responsible for the anticonvulsant/antiepileptic activity of sulfonamides and sulfamates (3). In fact, CA7 has been shown in a rat model to act as a developmental switch in GABAergic signaling in neurons (4, 5). The amino acid sequence of human CA7 is 95%, 94%, 89%, 76%, 72% and 70% identical to that of bovine, rat/mouse, canine, chicken, Xenopus, and zebrafish.

1. Montgomery, J.C. et al. (1991) Genomics 11:835.
2. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
3. Vullo, D. et al. (2005) Bioorg. Med. Chem. Lett. 15:971.
4. Ruusuvuori, E. et al. (2004) J. Neurosci. 24:2699.
5. Rivera, C. et al. (2005) J. Physiol. 562:27.

Bioinformatics

• Gene Symbol: CA7
• Uniprot:
  • NP_005173()