. Author manuscript; available in PMC: 2011 Oct 31.

Published in final edited form as: Anal Biochem. 2008 Jun 16;380(1):134–136. doi: 10.1016/j.ab.2008.05.041

Table 1.

Fluorescence steady state and decay parameters, FRET efficiencies and “proximity” corrections determined from steady state and time-resolved measurements of membranes containing mixtures of RRG380R mutants of FGFR3 peptide labeled with donor (Fl) and acceptor (Rh).

Sample	Relative Fluor. Intensity		Decay Parameters					E_SS	E_TR	E^Exp._Corr.	¹E^Theor._Corr.
Sample	I_D	I_DA	τ₁ ns	α₁	τ₂ ns	α₂	τ_α	E_SS	E_TR	E^Exp._Corr.	¹E^Theor._Corr.
0.125%RRG380R_Fl	1	-	2.9	0.49	4.7	0.51	3.8	-	-	-	-
0.04%RRG380R_Fl+0.04%RRG380R_Rh	1	0.91	2.9	0.54	4.7	0.46	3.7	0.09	0.03	0.06	0.06
0.15%RRG380R_Fl+0.15%RRG380R_Rh	1	0.75	2.4	0.49	4.3	0.51	3.4	0.25	0.11	0.14	0.14

from (4).

Note: I_D and I_DA are the steady-state relative donor intensities of samples containing only donor-labeled peptides and samples with both donor- and acceptor-labeled peptides, respectively. The amplitude-averaged lifetimes τ_α are defined by Eq.3. The experimentally corrected FRET efficiency (E^Exp._Corr.) is calculated as a difference between the total FRET efficiency obtained in a steady-state experiment (E_SS) and “proximity” FRET efficiency obtained in a time-resolved experiment (E_TR). The relative errors of E_SS, E_TR and E^Exp._Corr. were less than 1%, 15% and 10%, respectively. E^Theor._Corr. - theoretically corrected FRET efficiency (4)