Biological Activity
The activity was measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2. The specific activity is >150 pmoles/min/µg.Recombinant Human MMP-3 protein pro form needs to be activated with Chymotrypsin.Activation Protocol:1. Dilute MMP3 to 20µg/ml in the Assay Buffer: 50mM Tris, 10mM CaCl2, 150mM NaCl, 0.05% (w/v) and Brij 35, pH 7.5.2. Activate MMP3 by adding Chymotrypsin(Sigma, Catalog#C3142,1mg/ml stock in 1mM HCl) to a final concentration of 5µg/ml.3. Incubate at 37°C for 30 minutes.4. Stop activation with 2mM PMSF. Pre-warm the PMSF to 37°C prior to adding to sample.
