Recombinant Human Proprotein Convertase 2/PCSK2 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 NaOAc, 100 mM NaCl, 1 mM CaCl₂, 0.5% (w/v) Brij-35, pH 5.0
• Recombinant Human Proprotein Convertase 2/PCSK2 (rhPCSK2) (Catalog # 6018-SE)
• Substrate: L-PYROGlu-Arg-Thr-Lys-Arg-AMC (Catalog # ES013), 4 mM stock in deionized water
• F16 Black Maxisorp Plate (Nunc, Catalog # 475515
• Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent

1. Dilute rhPCSK2 to 0.4 µg/mL in Assay Buffer.
2. Dilute Substrate to 500 µM in Assay Buffer.
3. Start the reaction by mixing equal volumes of 0.4 µg/mL rhPCSK2 and 500 µM Substrate together and incubate at 37 °C for 30 minutes. For a Substrate Blank, mix Assay Buffer and Substrate together and incubate at 37 °C for 30 minutes.
4. Into a plate, load 100 µL of the reaction and Substrate Blank (in triplicate).
5. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in endpoint mode.
6. Calculate specific activity:

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

• rhPCSK2: 0.02 µg
• Substrate: 250 µM