Recombinant Human MMP-14/MT1-MMP 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.

• Activation Buffer: 50 mM Tris, 1 mM CaCl₂, 0.5% (w/v) Brij-35, pH 9.0
• Assay Buffer: 50 mM Tris, 3 mM CaCl₂, 1 µM ZnCl₂, pH 8.5
• Recombinant Human MMP‑14/MT1‑MMP (rhMMP-14) (Catalog # 918-MP)
• Recombinant Human Furin (rhFurin) (Catalog # 1503-SE)
• Substrate: MCA-Lys-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH₂ (Catalog # ES010) 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. Activate rhMMP-14 at 80 µg/mL with 1.72 µg/mL rhFurin in Activation Buffer.
2. Incubate at 37 °C for 1.5 hours.
3. Dilute activated rhMMP-14 to 1 ng/µL in Assay Buffer.
4. Dilute Substrate to 80 µM in Assay Buffer.
5. In a plate load 50 µL of 1 ng/µL rhMMP-14 to wells, and for a Substrate Blank load 50 µL of Assay Buffer.
6. Start the reaction by adding 50 µL of 80 µM Substrate to wells.
7. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
8. Calculate specific activity:

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