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Recombinant Human Carbonic Anhydrase IX Protein, CF

R&D Systems | Catalog # 2188-CA

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Carbonic Anhydrase IX Protein (2188-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

NS0

Accession Number

Q16790

Applications

Enzyme Activity
View all Carbonic Anhydrase IX/CA9 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Carbonic Anhydrase IX/CA9 protein
Pro59-Asp414, 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

Pro59

Predicted Molecular Mass

42 kDa

SDS-PAGE

53 kDa and 54 kDa, reducing conditions

Activity

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

Formulation, Preparation, and Storage

2188-CA
Formulation Lyophilized from a 0.2 μm filtered solution in Tris and NaCl.
Reconstitution Reconstitute at 100 μg/mL in sterile 25 mM Tris and 150 mM NaCl, pH 7.5.
Shipping The product is shipped at ambient temperature. 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 reconstitution.

Calculators

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 IX/CA9

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 (1-3). Topics in the 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. CA9, also known as membrane antigen MN and renal cell carcinoma (RCC)‑associated antigen G250, is a transmembrane enzyme expressed primarily in carcinoma cells. It is one of the best markers for hypoxia and for RCC (4, 5). rhCA9 corresponds to the extracellular portion of human CA9.

References

  1. Pastorek, J. et al. (1994) Oncogene 9:2877.
  2. Opavsky, R. et al. (1996) Genomics 33:480.
  3. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
  4. Kaluzova, M. et al. (2004) Mol. Cell Biol. 24:5757.
  5. Mukouyama, H. et al. (2004) Clin. Cancer Res. 10:1421.

Alternate Names

CA9, G250, MN, RCC

Entrez Gene IDs

768 (Human); 230099 (Mouse)

Gene Symbol

CA9

UniProt

Q16790

Additional Carbonic Anhydrase IX/CA9 Products

Product Documents for Recombinant Human Carbonic Anhydrase IX 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 IX Protein, CF

For research use only

Related Research Areas

Cancer Biomarkers

Citations for Recombinant Human Carbonic Anhydrase IX Protein, CF

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Protocols

View specific protocols for Recombinant Human Carbonic Anhydrase IX Protein, CF (2188-CA):

Materials
  • Assay Buffer: 12.5 mM Tris, 75 mM NaCl, pH 7.5
  • Recombinant Human Carbonic Anhydrase IX/CA9 (rhCA9) (Catalog # 2188-CA)
  • Substrate: 4-Nitrophenyl acetate (4-NPA) (Sigma, Catalog # N8130), 100 mM stock in acetone
  • 96-well Clear Plate (Costar, Catalog # 2592)
  • Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhCA9 to 20 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Load in plate, 50 µL of 20 ng/µL rhCA9, 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:
  • rhCA9: 1 µg
  • Substrate: 1 mM

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