Recombinant Human Carbonic Anhydrase VI Protein, CF Summary
Gln18-Asn308, with a C-terminal 10-His tag
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, NaCl, Brij and Glycerol.|
|Shipping||The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.|
|Stability & Storage:||Do not freeze.
- Assay Buffer: 12.5 mM Tris, 75 mM NaCl, pH 7.5
- Recombinant Human Carbonic Anhydrase VI (rhCA6) (Catalog # 2939-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 rhCA6 to 50 ng/µL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- Load in plate, 50 µL of 50 ng/µL rhCA6, 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.
- 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).
- rhCA6: 2.5 µg
- Substrate: 1 mM
Background: Carbonic Anhydrase VI
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 (1). Topics in a CA meeting (6th 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 VI, also known as gustin and salivary Carbonic Anhydrase, is a zinc-metalloprotein that constitutes about 3% of human parotid saliva protein (2, 3). It was decreased in patients with loss of taste and pathological changes in taste buds (4). It is also an elementary component of milk. It plays an important role in normal growth and development of the infant alimentary tract (5).
- Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
- Murakami, H. and Sly, W. S. (1987) J. Biol. Chem. 262:1382.
- Thatcher, B. J. et al. (1998) Biochem. Biophys. Res. Commun. 250:635.
- Hankin, R. I. et al. (1999) Am. J. Med. Sci. 318:380.
- Karhumaa, P. et al. (2001) Proc. Natl. Acad. Sci. USA. 98:11604.
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