Mujumdar R B, Ernst L A, Mujumdar S R, Waggoner A S
Department of Biological Sciences, Carnegie-Mellon University, Pittsburgh, Pennsylvania 15213.
Cytometry. 1989 Jan;10(1):11-9. doi: 10.1002/cyto.990100104.
New isothiocyanate derivatives of cyanine dyes were synthesized as fluorescent covalent labeling reagents for proteins and other biomolecules. These dyes have maximum absorbance in the red and near infrared regions of the spectrum, have high extinction coefficients and have adequate quantum yields. Incorporating two alkyl sulfonate groups in the dye structures increases their water solubility, which is beneficial for labeling biological molecules in aqueous solution. Reactivities of proteins with these new cyanines are similar to their reactivities with fluorescein isothiocyanate. These new labeling reagents are complementary to the fluorescein and rhodamine reagents, expanding the possibilities of multicolor analyses. Sheep anti-mouse-IgG antibody was labeled with a pentamethine cyanine dye (CY5.8-ITC) and used with a fluoresceinated antibody as a second reagent for detecting human T-cell subsets by flow cytometry.
合成了新型花青染料异硫氰酸酯衍生物，作为蛋白质和其他生物分子的荧光共价标记试剂。这些染料在光谱的红色和近红外区域具有最大吸收度，具有高消光系数和足够的量子产率。在染料结构中引入两个烷基磺酸盐基团可提高其水溶性，这有利于在水溶液中标记生物分子。蛋白质与这些新型花青的反应性与其与异硫氰酸荧光素的反应性相似。这些新型标记试剂与荧光素和罗丹明试剂互补，扩大了多色分析的可能性。用五甲川花青染料（CY5.8-ITC）标记羊抗鼠-IgG抗体，并与荧光素化抗体一起用作通过流式细胞术检测人T细胞亚群的第二种试剂。
