Fluorescence Polarisation (FP)

What is Fluorescence Polarisation?

Fluorescence polarisation (FP) is ideal to measure the binding of a small molecule to a much larger one. Unbound fluorescent tracers will depolarise light during emission resulting in low polarisation, because the unbound molecules tumble more rapidly than bound tracers. In contrast, bound fluorescent labelled molecules keep the polarisation orientation as their rotation is inhibited.

Measuring FP

To evaluate the polarization two emission measurements are needed: the first using a polarized emission filter parallel to the excitation filter (S-plane) and the second with a polarized emission filter perpendicular to the excitation filter (P-plane). Typical labels are FITC, BODIPY or TMR (TRITC).

Applications of FP

_CAMP assays
Immunoassays
Kinase assays
Receptor-ligand binding

Application Notes related to FP

Interaction between ER alpha and SCR-1 NR-2 through FP with Mithras Characterization of the interaction between recombinant estrogen receptor alpha and transcriptional coactivator SCR-1 NR-2 binding motif through Fluorescence Polarization study with Mithras LB 940

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