Recombinant Mouse Carbonic Anhydrase IV Protein, CF

R&D Systems | Catalog # 2414-CA

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Carbonic Anhydrase IV Protein (2414-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

Q64444

Applications

Enzyme Activity
View all Carbonic Anhydrase IV/CA4 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Carbonic Anhydrase IV/CA4 protein
Glu18-Ser277, with a C-terminal 10-His tag
Accession # Q64444

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

Glu18

Predicted Molecular Mass

31 kDa

SDS-PAGE

35-38 kDa, reducing conditions

Activity

Measured by its esterase activity.
The specific activity is >3 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

2414-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 IV/CA4

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. The deduced amino acid sequence of mouse CA4 consists of a signal peptide (residues 1-17), an ectodomain (residues 18-277) and a pro peptide (residues 278-305), which is removed in the mature form (2). The native enzyme is attached to the membrane by a GPI-anchor. Recombinant mouse CA4 corresponds to the ectodomain and exhibits the activity as described in Activity Assay Protocol.

References

  1. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
  2. Tamai, S. et al. (1996) Biochem. Genet. 34:31.

Alternate Names

CA4, RP17

Entrez Gene IDs

762 (Human); 12351 (Mouse)

Gene Symbol

CA4

UniProt

Q64444

Additional Carbonic Anhydrase IV/CA4 Products

Product Documents for Recombinant Mouse Carbonic Anhydrase IV 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 Mouse Carbonic Anhydrase IV Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Mouse Carbonic Anhydrase IV Protein, CF (2414-CA):

Materials
  • Assay Buffer: 12.5 mM Tris, 75 mM NaCl, pH 7.5
  • Recombinant Mouse Carbonic Anhydrase IV/CA4 (rmCA4) (Catalog # 2414-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 rmCA4 to 80 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Load 50 µL of 80 ng/µL rmCA4 into a 96 well clear well plate, and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing Assay Buffer in place of rmCA4.
  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:
  • rmCA4: 4 µg
  • Substrate: 1 mM

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