Friday, December 16, 2011

Focus on Fluorescence Polarization

Fluorescence Polarization is a technique that determines a fluorescent molecules ability to retain polarization and reflects the tumbling rate of molecules in solution. The sample will be excited with polarized light and the polarization value will be calculated from the parallel and perpendicular emission light. Large molecules, such as proteins in solution, rotate relatively slowly due to their size, and will emit polarized light. Conversely, fast rotation of smaller molecules will result in a depolarized signal. A low polarization value indicates that molecules move freely in solution. A high FP value then means that a larger molecular complex is present, which rotates slowly due to the size of the complex.

Simultaneous Dual Emission Detection is especially useful for Fluorescence Polarization assays. BMG LABTECH's Dual Emission Detection System utilizes two matched PMTs allowing the detection of two emitted wavelengths at the same time. The microplate has to be read only once and any signal variations due to differences in well volumes and concentrations or fluctuations in excitation energy are corrected. This allows for increased throughput, improved sensitivity and lower %CVs.

Fluorescence Polarization applications include binding assays, the Predictor hERG assay, Transcreener® ADP2 FP Assay, HitHunter™ IP3 Assay, and many more.

The Dept. of Pharmacology at the University of Cambridge  recently developed a versatile assay that uses fluorescence polarization and the PHERAstar  to quantify in medium-throughput assays the ligand recognition properties of theIP3-binding site. For more information, read their article here and visit our customer focus website:

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