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33645170011
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note
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The fluorescence quantum yield (QY) of fluorescein is not greatly lessened by attachment to a Con A surface, that is, QY of fluorescein - 0.95, QY of fluorescein-5-maleimide - 0.47, QY of FI-Con A 0.52 (see Figure S2 in the Supporting Information).
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20
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0016816432
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33645184802
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note
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7-Dimetylaminocomarin-4-acetic acid (see Figure S5a in the Supporting Information).
-
-
-
-
25
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-
33645169913
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note
-
As a standard method, the wavelength at 490 nm (energy donor) was used for quantitative analysis. However, the emission peak at 490 nm overlaps partially with the FI emission to make the precise analysis difficult. Thus, we normalized the spectra using the peak at 544 nm instead of 490 nm in this study.
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26
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33645171145
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note
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It is known that UV excitation is sometimes problematic because of the UV-induced strong photobleaching. However, we confirmed that the photobleaching of the emission of FI-Con A by 280 nm excitation scarcely occurred under our measurement conditions (see Figure S6 in the Supporting Information). Thus, it is considered that the luminescent ratio change in the titration experiments was not affected by this factor.
-
-
-
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27
-
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33645175852
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note
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III in these titrations, because the luminescence spectra of native Con A/Tb were never changed by the addition of Man-1 (see Figure S8 in the Supporting Information).
-
-
-
-
28
-
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33645176300
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-
note
-
When Fl-Con A was titrated against Man type saccharides, the fluorescence intensity was gradually lessened and saturated (e.g. Man-5, see Figure 59 in the Supporting Information). This spectral change is the same as that of IAEDANS-Con A, and thus the mechanism for the fluorescence change, that is, the fluorophore being pushed out by the sugar, as with IAEDANS-Con A, is suggested in this case.
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29
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0023665255
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32
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33645173765
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note
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The reaction was also monitored by MALDI-TOF MS. The time course was in agreement with that monitored by LRET.
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