Simultaneous detection of the subcellular localization of RNAs and proteins in cultured cells by combined multicolor RNA-FISH and IF.
Simultaneous detection of the subcellular localization of RNAs and proteins in cultured cells by combined multicolor RNA-FISH and IF.
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DOI：
10.1016/j.ymeth.2016.09.010
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发表时间：
2017-04-15
期刊：
影响因子：
--
通讯作者：
Tuschl T
中科院分区：
文献类型：
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作者：
Meyer C;Garzia A;Tuschl T
荧光原位杂交（FISH）和免疫荧光（IF）是检测培养细胞中核酸和蛋白质的灵敏技术。然而，这些技术很少一起应用，并且标准方案不容易兼容于同一标本上的顺序应用。在这里，我们提供了一个用户友好的一步一步的方案来执行多色RNA-FISH与IF结合，同时检测培养细胞中不同rna和蛋白质的亚细胞定位。我们通过分析暴露于各种应激条件下的细胞中rna和蛋白质亚细胞分布的变化来证明我们的方案的使用。
Fluorescence in situ hybridization (FISH) and immunofluorescence (IF) are sensitive techniques for detecting nucleic acids and proteins in cultured cells. However, these techniques are rarely applied together, and standard protocols are not readily compatible for sequential application on the same specimen. Here, we provide a user-friendly step-by-step protocol to perform multicolor RNA-FISH in combination with IF to simultaneously detect the subcellular localization of distinct RNAs and proteins in cultured cells. We demonstrate the use of our protocol by analyzing changes in the subcellular distribution of RNAs and proteins in cells exposed to a variety of stress conditions.