Cascade blue derivatives: water soluble, reactive, blue emission dyes evaluated as fluorescent labels and tracers.
Cascade blue derivatives: water soluble, reactive, blue emission dyes evaluated as fluorescent labels and tracers.
复制标题
级联蓝色衍生物：水溶性、反应性、蓝色发射染料，作为荧光标记和示踪剂进行评估。
DOI：
10.1016/0003-2697(91)90515-u
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发表时间：
1991
影响因子：
2.9
通讯作者：
Haugland,RP
中科院分区：
文献类型：
--
作者：
Whitaker,JE;Haugland,RP;Moore,PL;Hewitt,PC;Reese,M;Haugland,RP
制备并评价了基于芘氧基三磺酸（级联蓝）结构的荧光染料。染料含有与胺、硫醇、酸、醛和酮反应的官能团，形成具有广泛生物学意义的分子的共价键合的荧光衍生物。级联蓝染料中的反应性基团包括羧酸和活化酯、胺、酰肼、醇、光亲和试剂、丙烯酰胺和卤代乙酰胺。染料在374-378 nm和399-403 nm处有最大吸收，消光系数分别为1.9 × 104-2.4 × 104 m −1cm− 1和2.3 × 104-3.0 × 104 m −1cm−1。发射最大值范围为422-430 nm。荧光染料的光谱性质与荧光素的光谱性质完全不同，以允许同时使用两种染料，同时具有最小的光谱干扰。级联蓝衍生物具有比具有类似光谱性质的其它活性染料更窄的光谱带宽和更小的斯托克斯位移，不显示出对pH的明显敏感性，在水溶液中具有更高的溶解度，并且具有良好至优异的量子产率。制备了许多组织化学和生物学上有用的分子的Cascade Blue缀合物，包括葡聚糖、白蛋白、Fc受体结合蛋白、抗体、凝集素、膜受体结合蛋白和生物素结合蛋白，以及生物颗粒和细菌。二级和三级标记的级联蓝缀合物在人上皮细胞（HEp-2）核的间接免疫荧光染色中产生特异性荧光定位。
Fluorescent dyes based on the pyrenyloxytrisulfonic acid (Cascade Blue) structure were prepared and evaluated. The dyes contain functional groups that react with amines, thiols, acids, aldehydes, and ketones, forming covalently bonded, fluorescent derivatives of molecules with broad biological interest. Reactive groups in the Cascade Blue dyes include carboxylic acids and activated esters, amines, hydrazides, alcohols, photoaffinity reagents, acrylamides, and haloacetamides. The dyes exhibited absorption maxima at 374–378 nm and 399–403 nm, with extinction coefficients in the range of 1.9 × 104–2.4 × 104m−1cm−1and 2.3 × 104–3.0 × 104m−1cm−1, respectively. Emission maxima ranged from 422–430 nm. The spectral properties of the fluorescent dyes are sufficiently different from fluorescein to permit simultaneous use of both dyes with minimum spectral interference. The Cascade Blue derivatives have narrower spectral bandwidths and smaller Stokes' shifts than other reactive dyes with similar spectral properties, do not show appreciable sensitivity to pH, have higher solubilities in aqueous solution, and have good to excellent quantum yields. Cascade Blue conjugates of a number of histochemically and biologically useful molecules were prepared, including dextrans, albumins, Fcreceptor binding proteins, antibodies, lectins, membrane receptor binding proteins, and biotin binding proteins, as well as biological particles and bacteria. Cascade Blue conjugates of secondary and tertiary labels yielded specific fluorescence localization in the indirect immunofluorescent staining of human epithelial cell (HEp-2) nuclei.