| Assay Method Information | |
| | FRET assay |
| Description: | Dose response curves against Rv2613c with 1 as the substrate for derivatives 5 and 18. 8 nM of Rv2613c were preincubated in buffer (20 mM Tris-HCl (pH 8.0), 50 mM NaCl, 1 mM DTT, 0.2 mg/mL BSA, 5 mM NaH2PO4, 5 mM MgCl2) with indicated amounts of compound solved in DMSO at 25°C for 30 min. Substrate 1 (2 µM in Tris-HCl buffer pH 8.0) was added Measurement of the fluorescence at 570 nm was conducted every 3 min over 1 h. Turnover of probe 1 was calculated by normalization to the intensity of controls. The rate of probe 1 turnover was obtained by linear fitting of probe 1 turnover over time. % Inhibition was calculated relative to the controls. The IC50 was calculated by a non-linear curve fit/dose response curve in Origin. |
| Affinity data for this assay | |
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