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| | Fluorescence Polarization Assay |
| Description: | In the fluorescence polarization assay, the Bcl-XL protein is incubated with a fluorescein-tagged Bak-BH3 peptide. The Bcl-XL:Bak-BH3 peptide complex is then titrated with the test compounds, and the displacement of the Bak-BH3 peptide is measured as a function of decreasing polarized fluorescence. The fluorescence polarization values were determined using a Tecan GeniosPro plate reader using the excitation/emission wavelengths 485/535 nm. The binding affinities are reported as the inhibitory concentration of the titrant required to displace 50% of the Bak-BH3 peptide (IC50). |
| Affinity data for this assay | |
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