Recombinant Human Carbonic Anhydrase I Protein, CF

R&D Systems | Catalog # 2180-CA

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human Carbonic Anhydrase I Protein (2180-CA)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

E. coli

Accession Number

P00915

Applications

Enzyme Activity
View all Carbonic Anhydrase I/CA1 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Carbonic Anhydrase I/CA1 protein
Ala2-Phe261, with a C-terminal 10-His tag

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.

N-terminal Sequence Analysis

Ala2

Predicted Molecular Mass

30 kDa

SDS-PAGE

30 kDa, reducing conditions

Activity

Measured by its esterase activity.
The specific activity is >10 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

2180-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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Carbonic Anhydrase I/CA1

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 use of CAs as markers for tumor and hypoxia in clinic, as nutritional supplement in milk, and as a tool for CO2 removal and mosquito control in industry. CA1 is a cytosolic enzyme with the highest levels in erythrocytes and is a very early marker for erythroid differentiation (2). The activity of CA1 can also be measured by its ability to catalyze the reaction CO2 + H2O → HCO3- + H+, using a published method (3).

References

  1. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
  2. Sly, W.S. and P.Y. Hu (1995) Annu. Rev. Biochem. 64:375.
  3. Wilbur, K.M. and N.G. Anderson (1948) J. Biol. Chem. 176:147.

Alternate Names

CA1

Entrez Gene IDs

759 (Human)

Gene Symbol

CA1

UniProt

P00915

Additional Carbonic Anhydrase I/CA1 Products

Product Documents for Recombinant Human Carbonic Anhydrase I Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Carbonic Anhydrase I Protein, CF

For research use only

Citations for Recombinant Human Carbonic Anhydrase I Protein, CF

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Protocols

View specific protocols for Recombinant Human Carbonic Anhydrase I Protein, CF (2180-CA):

Materials
  • Assay Buffer: 12.5 mM Tris, 75 mM NaCl, pH 7.5
  • Recombinant Human Carbonic Anhydrase I/CA1 (rhCA1) (Catalog # 2180-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 rhCA1 to 100 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Load 50 µL of 100 ng/µL rhCA1 into a plate, and start the reaction by adding 50 µL of 2 mM Substrate.
  4. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
  5. Read at a wavelength of 400 nm (bottom read) in kinetic mode for 5 minutes.
  6. 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:
  • rhCA1: 5 µg
  • Substrate: 1 mM

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