| Assay Method Information | |
| | Inhibition Assay |
| Description: | MMP-8: 90 μL (29 ng) of human MMP-8 (Calbiochem #444229) was mixed with 10 μL of 10 mM APMA, and activated by reacting at 37° C. for 1 hour. 10 μL of the resultant enzyme solution was diluted to 90 μL with an assay buffer A. The mixture was added to 90 μL of a 20 μM fluorescent substrate (MOCAc-Pro-Leu-Gly-Leu-A2pr(Dnp)-Ala-Arg-NH2, Peptide Institute Inc., #3163-v) (SEQ ID NO: 3) prepared with an assay buffer B, and reacted at 37° C. for 5 hours. Enzyme activity was then determined by measuring with a fluorescence intensity meter (Labsystems, Fluoroskan Ascent) at an excitation wavelength of 320 nm and a measurement wavelength of 405 nm.From the enzyme activity in the presence or the absence of the test substance, the inhibition rate was determined, and the 50% inhibitiory concentration (IC50) of the test substance was calculated |
| Affinity data for this assay | |
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