Recombinant Human 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 Human Prolyl Oligopeptidase/PREP (rhPREP) (Catalog # 4308-SE)
• Substrate: Z-Gly-Pro-AMC (Bachem, Catalog # I-1145)
• F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
• Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent

1. Dilute rhPREP to 0.1 µg/mL in Assay Buffer.
2. Dilute Substrate to 100 µM in Assay Buffer.
3. Incubate at room temperature for 5 minutes.
4. Load 50 µL of 0.1 µg/mL of rhPREP into a plate, and start the reaction by adding 50 µL of 100 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
5. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
6. Calculate specific activity:

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

• rhPREP: 0.005 µg
• Substrate: 50 µM