The Fluorescence Polarisation (FP) assay measures changes in light polarization emitted by a small fluorescent tracer molecule appended to a peptide, nuclei acid or an antibody. It is widely used to monitor biomolecular interactions in solution and to provide a basis for direct and competitive interactions. The main advantage of FP is the possibility of performing as a homogenous assay, thereby making it suitable for higher throughput formats. When the fluorescently labelled molecule (tracer) is excited with plane-polarized light, it tumbles rapidly in the time between excitation and emission and therefore emits light that is depolarized. On the other hand, when tracer binds to much larger molecule like the target protein, its rotation is slower and the emission wavelength is polarized. A variation to this assay is Fluorescence anisotropy wherein the small molecule is fluorescently labelled and provides as a tracer.
- At o2h, we have successfully used FP assays to study the kinetics of target interactions with peptides, nucleic acids and small molecules in real time
- FP can be used for compound screening in higher throughput formats in microplates with a possibility of miniaturization and the derivation of EC50 values
- FP can be used as a primary screening platform or complimentary to other biophysical binding techniques such as Surface Plasmon Resonance (SPR), which is also extensively used at o2h for direct binding studies and FBDD
To know more about our biology services offering or to request our brochure, please reach out to us at discovery@o2h.com.
