Measuring membrane protein dynamics in neurons using fluorescence recovery after photobleach.
Measuring membrane protein dynamics in neurons using fluorescence recovery after photobleach.
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DOI：
10.1016/b978-0-12-391857-4.00006-9
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发表时间：
2012
影响因子：
--
通讯作者：
Henley, Jeremy M.
中科院分区：
文献类型：
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作者：
Gonzalez-Gonzalez, Inmaculada M.;Jaskolski, Frederic;Goldberg, Yves;Ashby, Michael C.;Henley, Jeremy M.
使用遗传编码的荧光标签如绿色荧光蛋白（GFP）作为报告分子来监测活细胞中的过程已经改变了细胞生物学。这些工具的一个主要应用是分析神经元中的蛋白质动力学。特别是，表面表达的荧光团标记的蛋白质的光漂白后的荧光恢复（FRAP）已经有助于解决关于神经元如何协调蛋白质的突触传递的突出问题。在这里，我们提供了一个概述的方法，设备和分析所需的执行，分析和解释这些实验。
The use of genetically encoded fluorescent tags such as green fluorescent protein (GFP) as reporters to monitor processes in living cells has transformed cell biology. One major application for these tools has been to analyze protein dynamics in neurons. In particular, fluorescence recovery after photobleach (FRAP) of surface expressed fluorophore-tagged proteins has been instrumental to addressing outstanding questions about how neurons orchestrate the synaptic delivery of proteins. Here, we provide an overview of the methodology, equipment, and analysis required to perform, analyze, and interpret these experiments.