Recombinant Mouse Carbonic Anhydrase IX Protein, CF

Catalog #: 2344-CA Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
2344-CA-010

R&D Systems Recombinant Proteins and Enzymes

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All Carbonic Anhydrase IX/CA9 Proteins and Enzymes

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Mouse Carbonic Anhydrase IX 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 >100 pmol/min/µg, as measured under the described conditions.

Source

Mouse myeloma cell line, NS0-derived mouse Carbonic Anhydrase IX/CA9 protein
Gln32-Asp390, with a C-terminal 10-His tag

Accession #

Q8VHB5

N-terminal Sequence
Analysis

Gln32

Predicted Molecular Mass

40 kDa

SDS-PAGE

42 kDa and 50 kDa, reducing conditions

Product Datasheets

2344-CA

Product Datasheet

COA

SDS

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.

2344-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: 25 mM Tris, 150 mM NaCl, pH 7.5
Recombinant Mouse Carbonic Anhydrase IX/CA9 (rmCA9) (Catalog # 2344-CA)
Substrate: 4-Nitrophenyl acetate (4-NPA) (Sigma, Catalog # N8130), 100 mM stock in Actone
96-well Clear Plate (Costar, Catalog # 92592)
Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent

Dilute rmCA9 to 4.0 ng/µL in Assay Buffer.
Dilute Substrate to 2 mM in Assay Buffer.
In a plate load 50 µL of 4.0 ng/µL rmCA9, and start the reaction by adding 50 µL of 2 mM Substrate to wells. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 2 mM Substrate.
Read at a wavelength of 400 nm (bottom read) in kinetic mode for 5 minutes.
Calculate specific activity:

Specific Activity (pmol/min/µg) =	Adjusted V_max* (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:

rmCA9: 0.2 µg
Substrate: 1 mM

Reconstitution Calculator

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). Topics in a CA meeting (6^th 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 (2, 3). Recombinant mouse CA9 corresponds to the extracellular portion of mouse CA9.

References

Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
Kaluzova, M. et al. (2004) Mol. Cell Biol. 24:5757.
Mukouyama, H. et al. (2004) Clin. Cancer Res. 10:1421.

Entrez Gene IDs

768 (Human); 230099 (Mouse)

Alternate Names

CA9; CAIX; CA-IX; Carbonate dehydratase IX; carbonic anhydrase 9; Carbonic Anhydrase IX; carbonic dehydratase; EC 4.2.1.1; G250; Membrane antigen MN; MN; P54/58N; PMW1; RCC; RCC-associated antigen G250; RCC-associated protein G250; Renal cell carcinoma-associated antigen G250