Recombinant Human Prostasin/Prss8 Protein, CF

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

• Assay Buffer: 50 mM Tris, 0.05% (w/v) Brij 35, pH 9.5
• Recombinant Human Prostasin/Prss8 (rhProstasin) (Catalog # 4599-SE)
• Substrate: BOC-Gln-Ala-Arg-AMC (Catalog # ES014), 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 rhProstasin to 20 µg/mL in Assay Buffer.
2. Dilute Substrate to 200 µM in Assay Buffer.
3. Load 50 µL of 20 µg/mL of rhProstasin into a well, and start the reaction by adding 50 µL of 200 µM Substrate. 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 (top read), respectively in kinetic mode for 5 minutes.
5. Calculate specific activity:

     *Adjusted for Substrate Blank

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

• rhProstasin:  1 µg
• Substrate: 100 µM