Recombinant Human Kallikrein 5 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: 0.1 M NaH₂PO₄, pH 8.0
• Recombinant Human Kallikrein 5 (rhKLK5) (Catalog # 1108-SE)
• Fluorogenic Peptide Substrate BOC-Val-Pro-Arg-AMC (Catalog # ES011), 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 rhKLK5 to 2 ng/µL in Assay Buffer.
2. Dilute Substrate to 200 µM in Assay Buffer.
3. Load 50 µL of the 2 ng/µL rhKLK5 into plate and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 200 µM Substrate without any rhKLK5.
4. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
5. Calculate specific activity:

     *Adjusted for Substrate Blank

     **Derived using calibration standard 7-Amino, 4-Methyl Coumarin (AMC) (Sigma, Catalog # A9891)

• rhKLK5: 0.1 µg
• Substrate: 100 µM