Principles of Fluorescence Spectroscopy

PRINCIPLES OF FLUORESCENCE SPECTROSCOPY 663
Figure 19.78. Fluorescence polarization immunoassay of creatine
kinase BB. Revised from [307].
that the polarization changes are smaller with the other
probes. This is because the lifetimes of these probes, the
fluorescein derivative IAF, and coumarin derivative CPM
are near 5 ns, while the lifetime of DANZA is near 15 ns.
The longer lifetime of the dansyl-labeled protein (DNS or
DANZA) allows more time for the protein to undergo rotational
diffusion. This limitation of FPIs to low-molecularweight
substances can be overcome by the use of long-lifetime
metal ligand probes, which is described in the following
chapter.
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