Matrix Metalloproteinase-3 Human Recombinant, HEK

Lyophilized powder at room temperature.

MMP-3 Human Recombinant produced in HEK293 cells is a proform of the Human MMP3 [Tyr18-Cys477 (Lys45Glu)] and fused with a ployhistide tag at the C-terminus, having an Mw of 52kDa. MMP-3 is purified by proprietary chromatographic techniques.
Source: HEK293 cells.
Physical Appearance and Stability: Store at 4C if entire vial will be used within 2-4 weeks. Store, frozen at -20C for longer periods of time.Avoid multiple freeze-thaw cycles.
Formulation and Purity: The MMP-3 is supplied as a 0.2um filtered solution in 20mM Tris-HCl, 150mM NaCl and 0.05% Brij35, pH 7.5. Greater than 95% as determined by SDS-PAGE.
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/ug.Recombinant Human MMP-3 protein pro form needs to be activated with Chymotrypsin.Activation Protocol:1. Dilute MMP3 to 20ug/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#C­ 3142, 1mg/ml stock in 1mM HCl) to a final concentration of 5ug/ml.3. Incubate at 37C for 30 minutes.4. Stop activation with 2mM PMSF. Pre-warm the PMSF to 37C prior to adding to sample.