| Assay Method Information | |
| | MMP-9 Assay |
| Description: | The MMP-9 assay was performed in 50 mM Tris-HCl buffer (pH 7.5, 10mM CaCl2, 150mM NaCl, 0.05% Brij-35). Then, 30ng Active MMP-9 was added to 50 μL of MMP-9 substrate solution (120 μM). After incubation for 25 min at 37°C, the reaction was stopped with 25 μL of 1% TFA. An aliquot of 25 μL internal standard solution was then added and the solution mixture was analysed by HPLC. Then 30ng MMP-9 was added to 30 μL of 50mM Tris-HCl buffer (pH 7.5, 10 mM CaCl2, 150 mM NaCl, 0.05%Brij-35) containing varying concentrations of GM6001 (0, 30, 60, 100, 300, 600, 1000, 2000, 4000 pM ) or inhibitor A (0, 15, 60, 105, 165, 900, 1500, 1800, 2100 nM). After incubation for 30 min at 37°C, 2 μL of 3 nM MMP-9 substrate was added. The final volume was adjusted to 50 μL and the final concentration of the peptide substrate was 120 μM. |
| Affinity data for this assay | |
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