Recombinant Human Carbonic Anhydrase VB Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human Carbonic Anhydrase VB Protein, CF Summary

Details of Functionality
Measured by its esterase activity. The specific activity is >150 pmol/min/µg, as measured under the described conditions.
Source
E. coli-derived human Carbonic Anhydrase VB/CA5B protein
Cys34-Pro317, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Cys34 & Ser35
Protein/Peptide Type
Recombinant Enzymes
Gene
CA5B
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

Theoretical MW
34 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
34 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, 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
  1. Dilute rhCA5B to 2 ng/µL in Assay Buffer.
  2. Dilute substrate to 2 mM in Assay Buffer.
  3. 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.
  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:
  • rhCA5B: 0.1 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 VB Protein, CF

  • CA5B
  • Carbonate dehydratase VB
  • carbonic anhydrase 5B, mitochondrial
  • Carbonic Anhydrase VB
  • carbonic anhydrase VB, mitochondrial
  • carbonic dehydratase
  • CAVB
  • CA-VB
  • EC 4.2.1.1
  • MGC39962

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 (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.

  1. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
  2. Fujikawa-Adachi, K. et al. (1999) J. Biol. Chem. 274:21228.
  3. Shah, G.N. et al. (2000) Proc. Natl. Acad. Sci. USA 97:1677.

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Bioinformatics

Gene Symbol CA5B
Entrez
Uniprot