Recombinant Cynomolgus Aminopeptidase PILS/ARTS1 Protein, CF

• Assay Buffer: 25 mM Tris, pH 8.0
• Recombinant Cynomolgus Monkey Aminopeptidase PILS/ARTS1 His-tag (cynoARTS1) (Catalog # 10237-ZN)
• Substrate: Leu-AMC (Bachem, Catalog # I-1240), 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 rcynoARTS1 to 1 µg/mL in Assay Buffer.
2. Dilute Substrate to 600 µM in Assay Buffer.
3. Load in plate 50 µL of 1 µg/mL rcynoARTS1, and start the reaction by adding 50 µL of 600 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 600 µM Substrate.
   
4. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode of 5 minutes.
5. Calculate specific activity:

*Adjusted for Substrate Blank
**Derived using calibration standard 7-amino, 4-Methyl Coumarin AMC (Sigma, Catalog # A9891)

• rcynoARTS1: 0.05 µg
• Substrate: 300 µM