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| | MBL Inhibitory Activity in Constructed E. coli Cells Expressing MBLs |
| Description: | a. Transfer 50 μl LB with 0.8 mM IPTG (0.4 mM final concentration in assay) to each wellb. Add 1 μl inhibitor/positive control (500/1000 μM final concentration in assay) to selected wellsc. Add 50 μl overnight culture (OD600=1) to each welld. Incubate in plate reader at 37° C. for 20 min with shakinge. Add 5 μl nitrocefin (3.2 mM) to selected wells with a multichannel pipettef. Incubate the plate in the plate reader for 3 hours, measuring nitrocefin hydrolysis at A482 every minute, with shaking in-betweeng. Measure endpoint at A600 to see how inhibitor alone effects bacterial growthh. Centrifuge the plate at 2800 rpm for 10 min, and transfer 50 μl to clean wellsi. Measure A482 endpoint. |
| Affinity data for this assay | |
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