Recombinant Human Airway Trypsin-like Protease/HAT, CF

• 6 months from date of receipt, -20 to -70 °C as supplied.
• 3 months, -20 to -70 °C under sterile conditions after reconstitution.

• Assay Buffer: 50 mM Tris, 0.05% (w/v) Brij-35, pH 9.5
• Recombinant Human Airway Trypsin‑like Protease/HAT (rhHAT) (Catalog # 2695-SE)
• Substrate: t-Butyloxycarbonyl Phe-Ser Arg 7-amino-4-methyl coumarin (Bachem, Catalog # I-1400), 10 mM stock in DMSO
• F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
• Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent

1. Dilute rhHAT to 0.02 ng/µL in Assay Buffer.
2. Dilute Substrate to 200 µM in Assay Buffer.
3. In a plate load 50 µL of 0.02 ng/µL rhHAT, and start the reaction by adding 50 µL of 200 µM Substrate to wells. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 200 µM Substrate.
4. Read at excitation and emission wavelengths of 380 nm and 460 nm, respectively in kinetic mode for 5 minutes.
5. Calculate specific activity:

     *Adjusted for Substrate Blank
     **Derived using calibration standard 7-amino, 4-Methyl Coumarin (Sigma, Catalog # A-9891).

• rhHAT: 0.001 µg
• Substrate: 100 µM