The copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction is increasingly used for detection of various macromolecules and metabolites in biological samples. Here, we present a detailed analysis of the CuAAC reaction conditions in cells and tissue sections. Using the optimized CuAAC conditions, we have devised a highly sensitive immunostaining technique, based on the tyramide signal amplification/catalyzed reporter deposition (TSA/CARD) method with a novel alkyne tyramide substrate. The described method offers improved detection threshold compared to conventional immunofluorescent staining and produces significantly lower non-specific background than TSA/CARD with fluorescent tyramides.

中文翻译：

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基于过氧化物酶介导的信号放大和点击化学的高灵敏度无背景免疫荧光染色的混合检测方法。

铜催化的叠氮化物-炔烃环加成反应（CuAAC）越来越多地用于检测生物样品中的各种大分子和代谢物。在这里，我们对细胞和组织切片中的CuAAC反应条件进行详细分析。使用优化的CuAAC条件，我们基于酪胺信号放大/催化报道分子沉积（TSA / CARD）方法和新型炔烃酪酰胺底物，设计了一种高灵敏度的免疫染色技术。与传统的免疫荧光染色相比，所描述的方法提供了更高的检测阈值，并且产生的非特异性背景显着低于带有荧光四酰胺的TSA / CARD。