| Assay Method Information | |
| | Inhibition Assay |
| Description: | MMP-3: Preparation of the MMP-3 for the activity assay: MMP-3 was provided as a stock solution (0.1 mg/mL). The enzyme was diluted in water to a final concentration of 0.005 mg/mL.MMP-3 peptide substrate was provided as a stock solution in DMSO (4.5 mM). For the assay, the substrate was diluted 1× reaction buffer to a concentration of 100 μM.Procedure: The enzymatic assays were performed in 96-well microtiter plate, which allowed simultaneous monitoring of multiple reactions. For each reaction, 84.6 μL of 1× reaction buffer (pH 7.6), 3.4 μL of MMP-3 (final concentration 7.7 nM) and 2 μL of the corresponding new compound were added to each well. A stock solution of new compound was prepared in DMSO (100 mM), and dilutions were prepared from this stock solution with DMSO. Finally, to the mixture contained in each well 10 μL of the substrate were added (final concentration 10 μM). The reaction was incubated for 4 hours at room temperature in an orbital shaker (90 rpm). |
| Affinity data for this assay | |
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