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Refining membrane penetration by a combination of steady-state and time-resolved depth-dependent fluorescence quenching

Author:
Kyrychenko, Alexander, Ladokhin, Alexey S.
Source:
Analytical biochemistry 2014 v.446 pp. 19-21
ISSN:
0003-2697
Subject:
fluorescence, fluorescent dyes, lipid bilayers, refining
Abstract:
Accurate determination of the depth of membrane penetration of a fluorescent probe, attached to a lipid, protein, or other macromolecule of interest, using depth-dependent quenching methodology is complicated by thermal motion in the lipid bilayer. Here, we suggest that a combination of steady-state and time-resolved measurements can be used to generate a static quenching profile that reduces the contribution from transverse diffusion occurring during the excited-state lifetime. This procedure results in narrower quenching profiles, compared with those obtained by traditional measurements, and thus improves precision in determination of the underlying depth distribution of the probe.
Agid:
5983809