TABLE 1.

Fluorescence Polarization Assay analysis of Bfl-1 proteins binding to BH3 peptides

Various concentrations of GST-Δα9-Bfl-1, GST-FL-Bfl-1, and GST-mut-FL-Bfl-1 proteins were incubated with 15 nm fluorescein isothiocyanate-conjugated peptides. Fluorescence polarization was measured after 10 min. The concentrations to achieve 50% maximal binding (EC₅₀) were calculated from the data and are presented in Table 1. These data are representative of three independent experiments.

Protein	Bim-BH3	Bak-BH3
Δα9-Bfl-1	66 nm	167 nm
FL-Bfl-1	<1 μm	<1 μm
mut-FL-Bfl-1	<1 μm	<1 μm