Recombinant Mouse Aminopeptidase N/CD13 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 Tris, pH 7.0
• Recombinant Mouse Aminopeptidase N/CD13 (rmCD13) (Catalog # 2335-ZN)
• Substrate: H-Ala-AMC (Bachem, Catalog # I-1410), 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 rmCD13 to 1 µg/mL in Assay Buffer.
2. Incubate at 37 °C for one hour (fully activates enzyme).
3. Dilute rmCD13 to 0.2 ng/µL in Assay Buffer.
4. Dilute Substrate to 200 µM in Assay Buffer.
5. Load 50 µL of 0.2 ng/µL rmCD13 in 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 200 µM Substrate.
6. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
7. Calculate specific activity:

     *Adjusted for Substrate Blank

     **Derived using calibration standard 7-Amino-4-Methyl-Courmarin (AMC) (Sigma, Catalog # A-9891).

• rmCD13 0.010 µg
• Substrate: 100 µM