Recombinant Human Carbonic Anhydrase VI Protein, CF

    
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  • Purity
    >90%, by SDS-PAGE under reducing conditions and visualized by silver stain
  • Endotoxin Level
    <1.0 EU per 1 μg of the protein by the LAL method.
  • Activity
    Measured by its esterase activity. The specific activity is >3 pmol/min/µg, as measured under the described conditions.
  • Source
    Mouse myeloma cell line, NS0-derived human Carbonic Anhydrase VI protein
    Gln18-Asn308, with a C-terminal 10-His tag
  • Accession #
  • N-terminal Sequence
    Analysis
    No results obtained: Gln18 predicted
  • Predicted Molecular Mass
    35 kDa
  • SDS-PAGE
    42 kDa, reducing conditions
Product Datasheets

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2939-CA
 
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.
  • 3 months from date of receipt, 2 to 8 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after opening.
 
Assay Procedure
Materials
  • 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
  1. Dilute rhCA6 to 50 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. 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.
  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:
  • 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).

  • References:
    1. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
    2. Murakami, H. and Sly, W. S. (1987) J. Biol. Chem. 262:1382.
    3. Thatcher, B. J. et al. (1998) Biochem. Biophys. Res. Commun. 250:635.
    4. Hankin, R. I. et al. (1999) Am. J. Med. Sci. 318:380.
    5. Karhumaa, P. et al. (2001) Proc. Natl. Acad. Sci. USA. 98:11604.
  • Entrez Gene IDs:
    765 (Human); 298657 (Rat)
  • Alternate Names:
    CA6; Carbonate dehydratase VI; carbonic anhydrase 6; Carbonic Anhydrase VI; carbonic anhydrase VIGUSTIN; CA-VI; EC 4.2.1.1; Gustin; MGC21256; Salivary carbonic anhydrase; Secreted carbonic anhydrase

FAQs

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Primary Antibodies
Description Application Cat# Citations Images  

His Tag Antibody

WB , Simple Western , Flow , AP , CyTOF-ready MAB050 32  
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His Tag HRP-conjugated Antibody

WB , Simple Western MAB050H 3  
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His Tag PE-conjugated Antibody

ICFlow IC050P 4  
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His Tag APC-conjugated Antibody

ICFlow IC050A  
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His Tag Antibody

WB , Flow , AP MAB050R 2  
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His Tag Biotinylated Antibody

WB BAM050 4  
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His Tag Alexa Fluor® 488-conjugated Antibody

ICFlow IC050G  
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His Tag Alexa Fluor® 700-conjugated Antibody

ICFlow IC050N  
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His Tag PerCP-conjugated Antibody

ICFlow IC050C  
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Protein Purification Kits
Description Application Cat# Citations Images  

Histidine-tagged Protein Purification Resin

IP999 1
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