Recombinant Mouse Cathepsin X/Z/P 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: 25 mM NaOAc, 5 mM DTT, pH 3.0
• Recombinant Mouse Cathepsin X/Z/P (rmCathepsin X/Z/P) (Catalog # 1033-CY)
• Fluorogenic Peptide Substrate V: MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys-(DNP)-OH (Catalog # ES005)
• F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
• Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent

1. Dilute rmCathepsin X/Z/P to 10 µg/mL in Assay Buffer.
2. Incubate at room temperature for 5 minutes to activate.
3. Dilute activated rmCathepsin X/Z/P to 0.4 ng/µL in Assay Buffer.
4. Dilute Substrate to 20 µM in Assay Buffer.
5. Load 50 µL of the 0.4 ng/µL rmCathepsin X/Z/P into a black well plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate without any rmCathepsin X/Z/P.
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)

• rmCathepsin X/Z/P: 0.02 µg
• Substrate: 10 µM