Binding and Diffusion
Performing confocal single molecule FRET experiments on the EI-FLEX allows for straightforward explorations of the range of conformations adopted by proteins under physiologically relevant conditions.
The smFRET and FCS capabilities of the EI-FLEX can both be employed to investigate the binding and dissociation of biomolecular complexes, the formation of aggregates and to perform studies of the competitive binding of small molecules – such as for small molecule drug screens.
The use of confocal single-molecule methods to study protein binding and dissociation offers several benefits:
- No surface immobilisation of the protein is required
- No flow-cell or complex experimental set-up is required
- Experiments can be performed under biologically relevant conditions
- Work directly with cell lysates without extensive purification
- Work at very low concentration (picomolar)
- Accurately measure very low dissociation constants (kDs)
Further Reading
