| Assay Method Information | |
| | Fluorescence Polarization Assay |
| Description: | The fluorescence polarization assay was performed as previously described [Jenkins et al., ACS Chem. Biol., 7:1170-1177]. In 384-well black Costar plates, LpxA or His6-LpxD were serially diluted while holding the fluorescent peptide at 20 nM in a final volume of 50 μl in 20mM HEPES pH 8.0 (0.01% DMSO). The wells were gently mixed and incubated at 30 °C in the dark for 15 min. Polarization was measured on a SpectraMax M5 plate reader in triplicate with readings taken at λex = 485 nm and λem = 525 nm. |
| Affinity data for this assay | |
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