Polarization with Fluorescein Label
November 27, 2012Download
Related Products: Gen5 for Detection
Gen5 version required: 1.0 or higher
Basis for the Assay:
Fluorescence Polarization is used extensively in research and screening laboratories. It can provide information on changes in molecular mobility found, for example, in receptor–ligand interactions, protein–DNA interactions, or membrane fluidity assays. It consists in two successive fluorescence measurements through a parallel and a perpendicular polarizer. The two results are automatically compared and polarization is reported. Any small fluorescent molecule can be used to design a polarization assay, and fluorescein is one of the most commonly used labels.
The protocol calls for a measurement using a 485/20 nm excitation filter and a 528/20 nm emission filter. Switching from the parallel to the perpendicular polarizer is automatic. The result of this Read step is two sets of raw data and calculation of polarization values based on the raw data.
The experiment file contains an example of simulated data.
The plate map contains 8 standards in duplicate, two blanks, and samples in duplicate.
The report includes a standard curve and statistics on the standard and sample replicates.