Recombinant Mouse Prolyl Oligopeptidase/PREP 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: 25 mM Tris, 250 mM NaCl, 2.5 mM DTT, pH 7.5
• Recombinant Mouse Prolyl Oligopeptidase/PREP (rmPREP) (Catalog # 6339-SE)
• Substrate: Z-GP-AMC (Bachem, Catalog # I-1145), 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 rmPREP to 0.1 ng/μL in Assay Buffer.
2. Dilute Substrate to 100 μM in Assay Buffer.
3. Load into a plate 50 μL of 0.1 ng/μL rmPREP, and start the reaction by adding 50 μL of 100 μM Substrate.
4. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 100 μM Substrate.
5. Read plate at excitation and emission wavelengths of 380 nm and 460 nm, respectively, in kinetic mode for 5 minutes.
6. Calculate specific activity:

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

• rmPREP: 0.005 μg
• Substrate: 50 μM