(a) Coating with FN
(b) Binding of EF-Tu to FN
Figure 5: Interaction of EF-Tu and fibronectin characterized by a PC-TIR sensor. Binding of an analyte to a reactant causes resonant wavelength shift that was measured throughout the process as a function of time. (a) Coating the sensor with fibronectin. (i) The sensor surface was calibrated with PBS and (ii) coated with fibronectin (FN at 200 μg/mL in a 200 μL volume of PBS at 25°C). Immobilization of FN onto the sensor caused resonant wavelength shift. (iii) The subsequent washes. (b) Binding of EF-Tu to FN. (I) Detection baseline of the FN functionalized sensor. (II) EF-Tu at 20 μg/mL (blue curve) or 50 μg/mL (red) and a negative control (a portion of proteoglycan 4) at 20 μg/mL (black) were incubated with FN at 25°C for the indicated times. (III) The sensor was washed.