Cyanine dye labeling reagents—carboxymethylindocyanine succinimidyl esters

Authors

  • Philip L. Southwick,

    1. Departments of Biological Sciences and Chemistry, and Center for Fluorescence Research, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213
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  • Lauren A. Ernst,

    1. Departments of Biological Sciences and Chemistry, and Center for Fluorescence Research, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213
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  • Erica W. Tauriello,

    1. Departments of Biological Sciences and Chemistry, and Center for Fluorescence Research, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213
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  • Stephen R. Parker,

    1. Departments of Biological Sciences and Chemistry, and Center for Fluorescence Research, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213
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  • Ratnakar B. Mujumdar,

    1. Departments of Biological Sciences and Chemistry, and Center for Fluorescence Research, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213
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  • Swati R. Mujumdar,

    1. Departments of Biological Sciences and Chemistry, and Center for Fluorescence Research, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213
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  • Hester A. Clever,

    1. Departments of Biological Sciences and Chemistry, and Center for Fluorescence Research, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213
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  • Dr. Alan S. Waggoner

    Corresponding author
    1. Departments of Biological Sciences and Chemistry, and Center for Fluorescence Research, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213
    • Center for Fluorescence Research, Carnegie Mellon University, Pittsburgh, PA 15213
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  • This work was supported in part by Grants NS 19353 and GM 34639 from the National Institutes of Health and in part by the Western Pennsylvania Advanced Technology Center through Pennsylvania's Ben Franklin Partnership Program.

Abstract

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.

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