| Assay Method Information | |
| | Measurement of Half Maximal Inhibitory Concentration (IC50) |
| Description: | In brief: precultures of P. aeruginosa PAO1 (5 mL) were grown in LB media for 13 hours at 37° C. and 220 rpm in 50 mL conical tubes (VWR International, PA). The cells were centrifuged for 5 min at 4,000 rpm and 4° C., washed two times and then diluted in buffer (100 mM KH2PO4 and 15 mM (NH4)2SO4 to an optical density at 600 nm (OD600) of 0.1. A small volume of compound stock solution (10 mM) was transferred to a microcentrifuge tube, initially diluted with DMSO to 20 μL, and then diluted to 1 mL with preculture cell suspension in defined media with OD600=0.0001, so the final DMSO concentration is 2%. The resultant cell suspension (200 μL) was transferred to a clear-bottom polystyrene 96-well plate (VWR International, PA) covered with a lid and incubated at 35° C. and 205 cpm for 24 h in a Synergy H1 microplate reader (Biotek Instruments Inc., Winooski, VT). |
| Affinity data for this assay | |
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