Figure 3. Thermodynamic binding curve of SYP and RCP.
(A) Values of FRET efficiency in each group of samples were detected. The blue line presents the binding of RCP with increasing concentrations of SYP fitting a curve of single-site binding equation. The red line presents the binding of RCP with increasing concentrations of YFP as reference. Data represent the means ± SEM of three independent measurements. (B) Redox status of RCP and purity of the FRET-based probes were measured by non-reducing SDS-PAGE. Lines shows purified oxidized RCP (through exposed to air for 2 hours), reduced RCP (through dialysis in DTT containing binding buffer), purified SYP, the unstained protein molecular weight marker (Thermo Scientific).
