School of Biosciences, University of Sheffield, Sheffield, UK.
Methods Mol Biol. 2022;2438:1-30. doi: 10.1007/978-1-0716-2035-9_1.
Here, we present a detailed protocol for fluorescence recovery after photobleaching (FRAP) to measure the dynamics of junctional populations of proteins in living tissue. Specifically, we describe how to perform FRAP in Drosophila pupal wings on fluorescently tagged core planar polarity proteins, which exhibit relatively slow junctional turnover. We provide a step-by-step practical guide to performing FRAP, and list a series of controls and optimizations to do before conducting a FRAP experiment. Finally, we describe how to present the FRAP data for publication.
在这里，我们呈现了一个详细的荧光漂白后恢复（FRAP）实验方案，用于测量活体组织中蛋白质连接群体的动力学。具体来说，我们描述了如何在果蝇蛹翅上对荧光标记的核心平面极性蛋白进行 FRAP，这些蛋白表现出相对较慢的连接周转率。我们提供了执行 FRAP 的分步实用指南，并列出了在进行 FRAP 实验之前要进行的一系列对照和优化。最后，我们描述了如何呈现 FRAP 数据以供发表。
