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  • 1
    ISSN: 1573-4994
    Keywords: Cyanine dyes ; energy transfer ; electron transfer ; Poisson distribution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract The cyanine dyes Cy5 and Cy5.5 are presented as a new long wavelength-excitable donor-acceptor dye pair for homogeneous fluoroimmunoassays. The deactivation pathways responsible for the quenching of the fluorescence of the antibody-bound donor are elucidated. Upon binding of the donor dye to the antibodies at low dye/protein ratios, its fluorescence quantum yield rises to unity. Higher dye/protein ratios lead to progressive aggregation of the dyes, which results in quenching of monomer fluorescence due to resonance energy transfer (RET) from the monomers to the nonfluorescent dimers. The dependence of the quenching efficiency on the labeling ratio is described quantitatively by assuming a Poisson distribution of the dyes over the antibodies. The maximum fluorescence intensity per antibody is obtained at a labeling ratio of 4. Upon formation of the antibody-antigen complex, electron transfer and RET to the antigen-bound acceptor dye occur. Steady-state and time-resolved fluorescence measurements reveal that approximately 50% of the donor quenching is due to RET, while the residual quenching effect is caused by the static quenching process.
    Type of Medium: Electronic Resource
    Library Location Call Number Volume/Issue/Year Availability
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  • 2
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Journal of High Resolution Chromatography 15 (1992), S. 659-664 
    ISSN: 0935-6304
    Keywords: Splitless injection, 100 μl volumes ; Injector insert, packing of ; Sample evaporation in vaporizing injector ; Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: If a sample evaporates by flash vaporization in an empty injector insert, the solute material is well mixed with the expanding solvent vapors and the maximum injection volume is determined by the requirement that no vapors must leave the vaporizing chamber. If evaporation occurs from a surface (e.g., of Tenax packing), however, the solvent evaporates first. The site of evaporation is cooled to the solvent's boiling point, and the cool island formed in the hot injector retains solutes of at least intermediate boiling point (visually observed for perylene). Solvent vapors, free from such solutes, may now expand backwards from the injector insert and leave through the septum purge exit. When solvent evaporation is complete, the site of evaporation warms up, causing the high boiling solutes to evaporate and to be carried into the column by the carrier gas. The technique somewhat resembles PTV injection, but is performed using a classical vaporizing injector.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    Library Location Call Number Volume/Issue/Year Availability
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