Recombinant Human Carbonic Anhydrase IV Protein, CF
Recombinant Human Carbonic Anhydrase IV Protein, CF Summary
Product Specifications
Ala19-Lys283, with a C-terminal 10-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
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.
|
Assay Procedure
- 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
- Dilute rhCA4 to 40 ng/µL in Assay Buffer.
- Dilute Substrate to 2.0 mM in Assay Buffer.
- 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.
- Cover and incubate at room temperature for 60 minutes in the dark.
- After incubation add 100 µL Absorbance Buffer and read plate immediately.
- Read (top read) absorbance in endpoint mode at 410 nm.
- 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).
- rhCA4: 2 μg
- Substrate: 0.5 mM
Reconstitution Calculator
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).
- Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
- Chandrashekar, J. et al. (2009) Science 326:443.
- Rebello, G. et al. (2004) Proc. Natl. Acad. Sci. USA 101:6617.
Citation for Recombinant Human Carbonic Anhydrase IV Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
1 Citation: Showing 1 - 1
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Ciliary Motility Decreased by a CO2/HCO3--Free Solution in Ciliated Human Nasal Epithelial Cells Having a pH Elevated by Carbonic Anhydrase IV
Authors: Okamoto, S;Yasuda, M;Kawaguchi, K;Yasuoka, K;Kikukawa, Y;Asano, S;Tsujii, T;Inoue, S;Amagase, K;Inui, TA;Hirano, S;Inui, T;Marunaka, Y;Nakahari, T;
International journal of molecular sciences
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot Control
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