Recombinant Human Pepsinogen C/PGC 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: 50 mM Sodium Citrate, 250 mM NaCl, pH 2.5
• Recombinant Human Pepsinogen C/PGC (rhPGC) (Catalog # 6186-AS)
• Substrate: MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(Dnp)-OH (Catalog # ES005), 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 rhPGC to 100 µg/mL in Assay Buffer.
2. Incubate 100 µg/mL rhPGC for 10 minutes at room temperature.
3. Dilute 100 µg/mL rhPGC to 0.5 µg/mL in Assay Buffer.
4. Dilute Substrate to 20 µM in Assay Buffer.
5. Load 50 µL of 0.5 µg/mL of rhPGC into a plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 20 µM Substrate.
6. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
7. Calculate specific activity:

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

• rhPGC: 0.025 µg
• Substrate: 10 µM