| Assay Method Information | |
| | Fluorescence Correlation Spectroscopy (FCS) |
| Description: | For the data shown, 25 nM BD-verapamil was added to various P-gp nanodisc concentrations in the presence of 1 μM empty nanodiscs, and 50 nM BD-vinblastine was added to P-gp nanodiscs in the presence of 0.3 μM empty nanodiscs. Experiments examining the competition between the fluorescent probes and drug substrates for binding were performed in the presence of a fixed concentration of P-gp nanodiscs(300 nM) and dye (as used for binding titrations). Drug stock solutions were created in DMSO, and the final DMSO concentration in FCS samples did not exceed 1% (v/v). Samples were incubated at room temperature (22.5 ± 0.5 °C) for 90 min before measurement; 1 min measurements were recorded for each sample 5−10 times. |
| Affinity data for this assay | |
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