Recombinant Mouse Carbonic Anhydrase IV Protein, CF

Catalog # Availability Size / Price Qty
2414-CA-010
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Mouse Carbonic Anhydrase IV 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 >3 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived mouse Carbonic Anhydrase IV/CA4 protein
Glu18-Ser277, with a C-terminal 10-His tag
Accession # Q64444
Accession #
N-terminal Sequence
Analysis
Glu18
Predicted Molecular Mass
31 kDa
SDS-PAGE
35-38 kDa, reducing conditions

Product Datasheets

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2414-CA

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.

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.

Assay Procedure

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
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 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.
Entrez Gene IDs
762 (Human); 12351 (Mouse)
Alternate Names
CA4; CAIV; CA-IV; Car4; Carbonate dehydratase IV; carbonic anhydrase 4; Carbonic Anhydrase IV; carbonic anhydrase IVRP17; carbonic dehydratase IV; EC 4.2.1.1; retinitis pigmentosa 17 (autosomal dominant); RP17

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