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Recombinant Human Carbonic Anhydrase VII Protein, CF

Catalog #: 3177-CA Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
3177-CA-050
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Recombinant Human Carbonic Anhydrase VII Protein, CF Summary

Product Specifications

Purity
>95%, 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 >20 pmol/min/µg, as measured under the described conditions.
Source
E. coli-derived human Carbonic Anhydrase VII/CA7 protein
Thr2-Ala264, with a C-terminal 10-His tag
Accession #
NP_005173
N-terminal Sequence
Analysis
Thr2
Predicted Molecular Mass
31 kDa
SDS-PAGE
31 kDa, reducing conditions

Product Datasheets

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3177-CA

Product Datasheet
COA
SDS

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

3177-CA

Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Assay Procedure

Materials
  • Assay Buffer: 12.5 mM Tris, 75 mM NaCl, pH 7.5
  • Recombinant Human Carbonic Anhydrase VII/CA7 (rhCA7) (Catalog # 3177-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 rhCA7 to 40 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Load in plate, 50 µL of 50 ng/µL rhCA7, 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:
  • rhCA7: 2 µg
  • Substrate: 1 mM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Carbonic Anhydrase VII/CA7

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 (2). 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 VII encoded by the CA7 gene is a cytosolic protein predominantly expressed in the salivary gland (1). CA7 may have additional tissue distributions and functions. For example, studies with CA inhibitors provide evidence that human CA7 is the CA isozyme responsible for the anticonvulsant/antiepileptic activity of sulfonamides and sulfamates (3). In fact, CA7 has been shown in a rat model to act as a developmental switch in GABAergic signaling in neurons (4, 5). The amino acid sequence of human CA7 is 95%, 94%, 89%, 76%, 72% and 70% identical to that of bovine, rat/mouse, canine, chicken, Xenopus, and zebrafish.

References
  1. Montgomery, J.C. et al. (1991) Genomics 11:835.
  2. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
  3. Vullo, D. et al. (2005) Bioorg. Med. Chem. Lett. 15:971.
  4. Ruusuvuori, E. et al. (2004) J. Neurosci. 24:2699.
  5. Rivera, C. et al. (2005) J. Physiol. 562:27.
Entrez Gene IDs
766 (Human)
Alternate Names
CA7; Carbonate dehydratase VII; carbonic anhydrase 7; Carbonic Anhydrase VII; carbonic anhydrase VIICAVII; carbonic dehydratase VII; CA-VII; EC 4.2.1.1

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