Recombinant Human Carbonic Anhydrase XIII Protein, CF

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

Recombinant Human Carbonic Anhydrase XIII Protein, CF Summary

• Details of Functionality: Measured by its esterase activity. The specific activity is >5 pmol/min/µg, as measured under the described conditions.
• Source: E. coli-derived human Carbonic Anhydrase XIII/CA13 protein
  Ser2-His262, with a C-terminal 10-His tag
• Accession #: Q8N1Q1
• N-terminal Sequence: Ser2
• Protein/Peptide Type: Recombinant Enzymes
• Gene: CA13
• 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: 33 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 reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in Tris and NaCl.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Reconstitution Instructions: Reconstitute at 1 mg/mL in sterile 25 mM Tris and 150 mM NaCl, pH 7.5.
• Assay Procedure:
  • Assay Buffer: 12.5 mM Tris, 75 mM NaCl, pH 7.5
  • Recombinant Human Carbonic Anhydrase XIII/CA13 (rhCA13) (Catalog # 2194-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 rhCA13 to 100 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2.0 mM in Assay Buffer.
  3. In a plate load 50 µL of 100 ng/µL rhCA13, and include a Substrate Blank containing 50 µL Assay Buffer.
  4. Start the reaction by adding 50 µL of 2.0 mM Substrate to wells containing rhCA13 and Assay Buffer.
  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:
  • rhCA13: 5 µg
  • Substrate: 1.0 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 XIII Protein, CF

• CA13
• carbonic anhydrase 13
• Carbonic Anhydrase XIII
• carbonic anhydrase XIIICarbonate dehydratase XIII
• CAXIII
• CA-XIII
• EC 4.2.1.1
• FLJ37995
• MGC59868

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. CA13 is a cytosolic enzyme with a unique and widespread distribution pattern as compared to the other cytosolic CAs, indicating that it may play important physiological roles in several organs (2).

1. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
2. Lehtonen, J. et al. (2003) J. Biol. Chem. 279: 2719.

Bioinformatics

• Gene Symbol: CA13
• Uniprot:
  • Q8N1Q1()