Recombinant Mouse Active Trypsin 3/PRSS3 Protein, CF

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

• Assay Buffer: 50 mM Tris, 10 mM CaCl₂, 150 mM NaCl, 0.05% (w/v) Brij35, pH 8.0
• Recombinant Mouse Active Trypsin 3/PRSS3 (rmTrypsin 3) (Catalog # 7235-SE)
• Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys-(DNP)-NH₂(Catalog # ES002), 2 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 rmTrypsin 3 to 0.007 ng/μL in Assay Buffer.
2. Dilute Substrate to 20 μM in Assay Buffer.
3. Load into a plate 50 μL of 0.007 ng/μL rmTrypsin 3, and start the reaction by adding 50 μL of 20 μM Substrate. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 20 μM Substrate.
4. Read plate at excitation and emission wavelengths of 320 nm and 405 nm, respectively, in kinetic mode for 5 minutes.
5. Calculate specific activity:

     *Adjusted for Substrate Blank
     **Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

• rmTrypsin 3: 0.35 ng (0.00035 μg)
• Substrate: 10 μM