Recombinant Human Carbonic Anhydrase VI Protein, CF

• 3 months from date of receipt, 2 to 8 °C as supplied.
• 1 month, 2 to 8 °C under sterile conditions after opening.

• Assay Buffer: 12.5 mM Tris, 75 mM NaCl, pH 7.5
• Recombinant Human Carbonic Anhydrase VI (rhCA6) (Catalog # 2939-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 rhCA6 to 50 ng/µL in Assay Buffer.
2. Dilute Substrate to 2 mM in Assay Buffer.
3. Load in plate, 50 µL of 50 ng/µL rhCA6, and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 2 mM Substrate.
4. Read at a wavelength of 400 nm (bottom read) in kinetic mode for 5 minutes.
5. Calculate specific activity:

     *Adjusted for Substrate Blank
     **Derived using calibration standard 4-Nitrophenol (Sigma, Catalog # 241326).

• rhCA6: 2.5 µg
• Substrate: 1 mM