Recombinant Human Carbonic Anhydrase VB Protein, CF
Recombinant Human Carbonic Anhydrase VB Protein, CF Summary
Product Specifications
Cys34-Pro317, with a C-terminal 10-His tag
Analysis
Product Datasheets
Carrier Free
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.
3176-CA
| 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 Procedure
- Assay Buffer: 12.5 mM Tris, 75 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5
- Recombinant Human Carbonic Anhydrase VB (rhCA5B) (Catalog # 3176-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 rhCA5B to 2 ng/µL in Assay Buffer.
- Dilute substrate to 2 mM in Assay Buffer.
- Load in plate, 50 µL of 2 ng/µL rhCA5B, and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing 50 µL of 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).
- rhCA5B: 0.1 µg
- Substrate: 1 mM
Reconstitution Calculator
Background: Carbonic Anhydrase VB/CA5B
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 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 VB encoded by the CA5B gene is a mitochondrial protein. In comparison with another mitochondrial CA (CA5A), CA5B has different tissue distribution and chromosomal location (2, 3). The amino acid sequence of human CA5B (residues 34 to 317) is 94%, 93%, 92%, and 74% identical to that of canine, mouse, bovine/rat, and chicken.
- Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
- Fujikawa-Adachi, K. et al. (1999) J. Biol. Chem. 274:21228.
- Shah, G.N. et al. (2000) Proc. Natl. Acad. Sci. USA 97:1677.
FAQs
No product specific FAQs exist for this product, however you may
View all Proteins and Enzyme FAQsReviews for Recombinant Human Carbonic Anhydrase VB Protein, CF
There are currently no reviews for this product. Be the first to review Recombinant Human Carbonic Anhydrase VB Protein, CF and earn rewards!
Have you used Recombinant Human Carbonic Anhydrase VB Protein, CF?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥2500 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image
