Galloylated catechins, as the main secondary metabolites in the tea plant, including (-)-epigallocatechin-3-gallate and (-)-epicatechin-3-gallate, comprise approximately three-quarters of all the tea plant catechins and have stronger effects than non-galloylated catechins, both on the product quality in tea processing and the pharmacological efficacy to human beings. The subcellular localization of galloylated catechins has been the primary focus of studies that assess biosynthesis and physiological functions. Classical histochemical localization staining reagents can not specifically detect galloylated catechins; thus, their subcellular localization remains controversial. In the present study, we generated a monoclonal antibody (mAb) against galloylated catechins, which can be used for the subcellular localization of galloylated catechins in the tea plant by immunohistochemistry. Direct ELISA and ForteBio Octet Red 96 System assay indicated the mAb could recognize the galloylated catechins with high specificities and affinities. In addition, tea bud was ascertained as the optimal tissue for freezing microtomic sections for immunohistochemistry. What's more, the high quality mAbs which exhibited excellent binding capability to galloylated catechins were utilized for the visualization of them via immunohistochemistry. Our findings demonstrated that vacuoles were the primary sites of localization of galloylated catechins at the subcellular level.

中文翻译：

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通过免疫组织化学评估了茶多酚中的儿茶素的亚细胞定位[Camellia sinensis（L.）O. Kuntze]。

Galloylated catechins作为茶树中的主要次要代谢产物，包括（-）-epigallocatechin-3-gallate和（-）-epicatechin-3-gallate，约占所有茶树儿茶素的四分之三，其作用强于非铝化儿茶素，无论是在茶叶加工中的产品质量还是对人类的药理功效。没食子酸儿茶素的亚细胞定位一直是评估生物合成和生理功能的研究的主要重点。经典的组织化学定位染色剂不能特异性检测没食子酸儿茶素。因此，它们的亚细胞定位仍然存在争议。在本研究中，我们产生了针对没食子儿茶素的单克隆抗体（mAb），通过免疫组织化学可将其用于茶树中没食子酸儿茶素的亚细胞定位。直接ELISA和ForteBio Octet Red 96系统测定表明，mAb可以识别具有高特异性和亲和力的没食子酸儿茶素。另外，确定茶芽是冷冻用于免疫组织化学的切片切片的最佳组织。此外，通过与免疫组化技术的可视化，显示出了与没食子酸儿茶素具有极佳结合能力的高质量mAb。我们的发现表明，液泡是在亚细胞水平上定位没食子儿茶素的主要部位。直接ELISA和ForteBio Octet Red 96系统测定表明，mAb可以识别具有高特异性和亲和力的没食子酸儿茶素。另外，确定茶芽是冷冻用于免疫组织化学的切片切片的最佳组织。此外，通过与免疫组化技术的可视化，显示出了与没食子酸儿茶素具有极佳结合能力的高质量mAb。我们的发现表明，液泡是在亚细胞水平上定位没食子儿茶素的主要部位。直接ELISA和ForteBio Octet Red 96系统测定表明，mAb可以识别具有高特异性和亲和力的没食子酸儿茶素。另外，确定茶芽是冷冻用于免疫组织化学的切片切片的最佳组织。此外，通过与免疫组化技术的可视化，显示出了与没食子酸儿茶素具有极佳结合能力的高质量mAb。我们的发现表明，液泡是在亚细胞水平上定位没食子儿茶素的主要部位。通过与免疫组化的可视化，显示了与没食子儿茶素具有极好的结合能力的高质量mAb。我们的发现表明，液泡是在亚细胞水平上定位没食子儿茶素的主要部位。通过与免疫组化的可视化，显示了与没食子儿茶素具有极好的结合能力的高质量mAb。我们的发现表明，液泡是在亚细胞水平上定位没食子儿茶素的主要部位。