| Assay Method Information | |
| | MMP12 Inhibition Assay |
| Description: | The synthesized compounds were assayed for the inhibitory activities against MMP-12 in 96-well microtiter plates using thiopeptolide as the substrate. MMP-12 was diluted in Assay buffer (50 mM Tris pH7.5, 0.05% Brij-35, 10 mM CaCl2, 1 mM DTNB) to a concentration of 0.007 U/µL. The MMP-12 solution (44 µL) was premixed with 1 µL of inhibitors dissolved in DMSO in a 96-well plate, and the mixture was incubated for 20 min at room temperature. Then, 5 µL of 1 mM MMP chromogenic substrate (thiopeptolide) was added to the mixture, and the reaction mixture was incubated for 30-80 min at 37 °C. Reaction was terminated by adding 7 µL of 0.2 M EDTA (pH 8.0). The intensity of the color developed by the digested substrate was measuredat 405 nm. |
| Affinity data for this assay | |
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