Binding and diffusion

The smFRET and FCS capabilities of our EI-FLEX can enable investigations into the binding and dissociation of biomolecular complexes and many more applications.

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

Binding and diffusion chart Fig 1. Overlays of the FCS correlation curves for free dye, a dye labelled antibody and a complex i.e. bound protein or vesicle containing the labelled antibody. As size increases so does diffusion time, which is the basis of the FCS technique.

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