| Assay Method Information | |
| | Fluorescence Polarization Assay |
| Description: | Fluorescence polarization experiments were conducted on a Photon Technology International instrument using a 0.3 cm path length cuvette. Spectra were measured at 25 deg C using 10.0 nm slit widths. Excitation at 495 nm was used for the fluorescein-containing peptide Fl-Bak and the emission maximum at 535 nm was monitored. Polarization measurements were recorded upon titration of inhibitors at varying concentrations into a solution of Fl-Bak and Bcl-xL. Regression analysis was carried out using SigmaPlot 2001 (Systat Co.) ligand binding macro module. Experimental data were fitted to determine the IC50 values, which can be related to the known affinity of the 16-mer Bak peptide (Kd =120 nM) to acquire the inhibitory constant Ki. |
| Affinity data for this assay | |
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