Recombinant Human Carbonic Anhydrase IV Protein, CF

R&D Systems | Catalog # 2186-CA

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Carbonic Anhydrase IV Protein (2186-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

P22748

Applications

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

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Carbonic Anhydrase IV/CA4 protein
Ala19-Lys283, 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

Ala19

Predicted Molecular Mass

32 kDa

SDS-PAGE

33 kDa, reducing conditions

Activity

Measured by its esterase activity.
The specific activity is >2 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

2186-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 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. CA4 is a GPI‑anchored membrane enzyme expressed on the luminal surfaces of pulmonary (and certain other) capillaries and of proximal renal tubules. It functions as the principal CO2 taste sensor (2). In addition, a genetic mutation (Arg 14 to Trp in the signal peptide) of CA4 was found to cosegregate with the RP17 form of retinitis pigmentosa in two large families and was not found in 36 unaffected family members or 100 controls (3).

References

  1. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
  2. Chandrashekar, J. et al. (2009) Science 326:443.
  3. Rebello, G. et al. (2004) Proc. Natl. Acad. Sci. USA 101:6617.

Alternate Names

CA4, RP17

Entrez Gene IDs

762 (Human); 12351 (Mouse)

Gene Symbol

CA4

UniProt

P22748

Additional Carbonic Anhydrase IV/CA4 Products

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

For research use only

Citations for Recombinant Human Carbonic Anhydrase IV Protein, CF

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Protocols

View specific protocols for Recombinant Human Carbonic Anhydrase IV Protein, CF (2186-CA):

Materials
  • Assay Buffer: 12.5 mM Tris, 75 mM NaCl, pH 7.5
  • Absorbance Buffer: 12.5 mM Tris, pH 8.0
  • Recombinant Human Carbonic Carbonic Anhydrase IV/CA4 (rhCA4) (Catalog # 2186-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 rhCA4 to 40 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2.0 mM in Assay Buffer.
  3. Load 50 µL of 40 ng/µL rhCA4 into plate and start the reaction by adding 50 µL of 2.0 mM Substrate. As a Substrate Blank combine 50 µL Assay Buffer with 50 µL Substrate.
  4. Cover and incubate at room temperature for 60 minutes in the dark.
  5. After incubation add 100 µL Absorbance Buffer and read plate immediately.
  6. Read (top read) absorbance in endpoint mode at 410 nm.
  7. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Abs* (OD) x Conversion Factor** (pmol/OD)
Incubation time (min) x amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard p-Nitrophenol (Sigma-Aldrich, Catalog # 241326).

Per Well:
  • rhCA4: 2 μg
  • Substrate: 0.5 mM

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