| Assay Method Information | |
| | Inhibition of Gelatinase E Assay |
| Description: | Gelatinase E (Gel E) was purified from bacterial culture supernatant from E. faecalis. E. faecalis was cultured aerobically in Todd Hewitt Broth overnight at 37 C. Nucleic acid was precipitated with 0.9% protamine solution, followed by protein precipitation with ammonium sulfate. Resuspended protein pellet was further subjected to purification using FPLC (phenyl Sepharose column). Fractions with gelatinase activity as determined by casein agar assay were pooled and further concentrated. Different concentrations of test compound were incubated with purified GelE and FRET-based peptide substrate (390 MMP FRET Substrate 1; Anaspec AS-27077) in assay buffer at room temperature for 30 minutes. The fluorescence signal was determined by a plate reader.Various compounds in Table 10 inhibited GelE with an IC50 of greater than 200 μM. Compound 62 showed high levels of inhibition, with an IC50 of about 16.42 μM. Compound A also showed high levels of GelE inhibition. |
| Affinity data for this assay | |
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