Recombinant Human Proprotein Convertase 1/PCSK1 Protein, CF

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

• Assay Buffer: 25 mM MES, 5 mM CaCl₂, 1% (w/v) Brij-35, pH 6.0.
• Recombinant Human Proprotein Convertase 1/PCSK1 (rhPCSK1) (Catalog # 2810-SE)
• Substrate: L-PyroGlu-Arg-Thr-Lys-Arg-AMC (pERTKR-AMC) (Catalog # ES013)
• F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
• Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent

1. Dilute rhPCSK1 to 4 µg/mL in Assay Buffer.
2. Dilute Substrate to 200 µM in Assay Buffer.
3. Load 50 µL of 4 µg/mL of rhPCSK1 into a plate, 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 Substrate.
4. Incubate at 37 °C for 1 hour.
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 # A9891).

• rhPCSK1: 0.200 µg
• Substrate: 100 µM