Recombinant Mouse/Rat MMP-2 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/Rat MMP-2 (rm/rMMP-2) (Catalog # 924-MP)
• p-aminophenylmercuric acetate (APMA) (Sigma, Catalog # A-9563), 100 mM Stock in DMSO
• Substrate: MCA-Pro-Leu-Gly-Leu-(DPA)-Ala-Arg-NH₂ (Catalog # ES001)
• F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
• Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent

1. Dilute rm/rMMP-2 to 40 µg/mL with 1 mM APMA in Assay Buffer.
2. Incubate at 37 °C for 2 hours to activate rm/rMMP-2.
3. Dilute activated rm/rMMP-2 to 0.2 ng/µL in Assay Buffer.
4. Dilute Substrate to 80 µM in Assay Buffer.
5. Load 50 µL of 0.2 ng/µL rm/rMMP-2 in a plate, and start the reaction by adding 50 µL of 80 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 80 µM Substrate.
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).

• rm/rMMP-2: 0.010 µg
• Substrate: 40 µM