Recombinant Mouse Carbonic Anhydrase XII Protein, CF
Recombinant Mouse Carbonic Anhydrase XII Protein, CF Summary
Ala25-Ser301, with a C-terminal 10-His tag
Accession # Q8CI85
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|Formulation||Supplied as a 0.2 μm filtered solution in Tris and NaCl.|
|Shipping||The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.|
|Stability & Storage:||Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 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
- Dilute rmCA12 to 8 ng/µL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- In a plate load 50 µL of 8 ng/µL rmCA12 and include a Substrate Blank containing 50 µL Assay Buffer.
- Start the reaction by adding 50 µL of 2 mM Substrate to wells.
- Read at a wavelength of 400 nm (bottom read) in kinetic mode for 5 minutes.
- 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).
- rmCA12: 0.40 µg
- Substrate: 1 mM
Background: Carbonic Anhydrase XII/CA12
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 (6th 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.
- Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
- Strausberg, R.L. et al. (2002) Proc. Natl. Acad. Sci. USA 99:16899.
- Okazaki, Y. et al. (2002) Nature 420:563.
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