Southwick P L, Ernst L A, Tauriello E W, Parker S R, Mujumdar R B, Mujumdar S R, Clever H A, Waggoner A S
Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213.
Cytometry. 1990;11(3):418-30. doi: 10.1002/cyto.990110313.
Ten carboxymethylindocyanine dyes which form 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 previously (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.
已经合成了十种羧甲基吲哚菁染料，它们构成了一系列新型荧光探针的基础，并被转化为琥珀酰亚胺活性酯，用于蛋白质或其他含氨基物质的荧光标记。该系列成员的最大荧光发射波长范围为575至780nm。已获得亲水性、水溶性试剂，这些试剂产生的标记抗体几乎没有形成沉淀的倾向。所达到的荧光强度高于先前描述的用异硫氰酸菁染料标记所产生的荧光强度（Mujumdar等人：《细胞计数》10：11 - 19，1989）。这些试剂的实用性已在抗体标记中得到证明，可用于哺乳动物细胞内部结构的双色免疫荧光成像以及双色流式细胞术实验。建议使用根据染料样品的定量吸收数据计算出的发色团当量重量（W/Ceq）值，以辅助制定标记程序。
