Recombinant Mouse MMP-7 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, 10 mM CaCl₂, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
• Recombinant Mouse MMP-7 (rmMMP-7) (Catalog # 2967-MP)
• p-aminophenylmercuric acetate (APMA) (Sigma, Catalog # A-9563), 100 mM stock in DMSO
• Substrate: MCA-Lys-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH₂, (Catalog # ES010)
• F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
• Fluorescent Plate Reader (Model: Gemini EM by Molecular Devices) or equivalent

1. Dilute rmMMP-7 to 100 μg/mL in Assay Buffer.
2. Activate rmMMP-7 by adding APMA to a final concentration of 1 mM.
3. Incubate at 37 °C for 1 hour.
4. Dilute activated rmMMP-7 to 0.4 ng/μL in Assay Buffer.
5. Dilute Substrate to 120 μM in Assay Buffer.
6. In a plate, load 50 μL of 0.4 ng/μL rmMMP-7 and start the reaction by adding 50 μL of 120 μM Substrate. Include a Substrate Blank containing 50 μL Assay Buffer and 50 μL of 120 μM Substrate.
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).

• rmMMP-7: 0.020 μg
• Substrate: 60 μM