Open AccessCyanine dye labeling reagents—carboxymethylindocyanine succinimidyl esters
Author(s) -
Southwick Philip L.,
Ernst Lauren A.,
Tauriello Erica W.,
Parker Stephen R.,
Mujumdar Ratnakar B.,
Mujumdar Swati R.,
Clever Hester A.,
Waggoner Alan S.
Publication year - 1990
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/cyto.990110313
Subject(s) - cyanine , reagent , fluorescence , chemistry , chromophore , flow cytometry , yield (engineering) , absorption (acoustics) , chromatography , photochemistry , organic chemistry , microbiology and biotechnology , materials science , biology , physics , quantum mechanics , metallurgy , composite material
Ten carboxymethylindocyanine dyes which from the basis of a new series of fluorescent probes have been synthesized and converted into succinimidyl active esters for fluorescent labeling of proteins or other amino‐containing substances. Fluorescence emission maxima for members of the series range from 575 to 780 nm. Hydrophilic, water‐soluble reagents have been obtained which yield labeled antibodies with little tendency to form precipitates. The fluorescence intensities achieved are higher than those produced by labeling with the cyanine isothiocyanates described previosly (Mujumdar et al.: Cytometry 10:11–19, 1989). The utility of these reagents has been demonstrated in antibody labeling for two‐color immunofluorescent imaging of internal structures in a mammalian cell and for two‐color flow‐cytometry experiments. The use of values of chromophore‐equivalent weight (W/Ceq), calculated from quantitative absorption data on dye samples, is proposed as an aid in formulating labeling procedures.