利用基于红色荧光蛋白的 FRET 对进行 Rho GTPase 活性的 FRET 成像。
FRET Imaging of Rho GTPase Activity with Red Fluorescent Protein-Based FRET Pairs.
机构信息
Department of Biological Chemistry, Medical Scientist Training Program, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA.
Guangdong Provincial Key Laboratory of Biomedical Optical Imaging Technology & Center for Biomedical Optics and Molecular Imaging, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
出版信息
Methods Mol Biol. 2022;2438:31-43. doi: 10.1007/978-1-0716-2035-9_2.
With the development of fluorescent proteins (FPs) and advanced optical microscopy techniques, Förster or fluorescence resonance energy transfer (FRET) has become a powerful tool for real-time noninvasive visualization of a variety of biological processes, including kinase activities, with high spatiotemporal resolution in living cells and organisms. FRET can be detected in appropriately configured microscopes as changes in fluorescence intensity, lifetime, and anisotropy. Here, we describe the preparation of samples expressing FP-based FRET sensors for RhoA kinase, intensity- and lifetime-based FRET imaging, and postimaging data analysis.
随着荧光蛋白（FPs）和先进的光学显微镜技术的发展，Förster 或荧光共振能量转移（FRET）已经成为一种强大的工具，可实时、非侵入式地可视化各种生物过程，包括激酶活性，具有高时空分辨率的活细胞和生物体。FRET 可以在适当配置的显微镜中作为荧光强度、寿命和各向异性的变化来检测。在这里，我们描述了用于 RhoA 激酶的基于 FP 的 FRET 传感器的样品制备、基于强度和寿命的 FRET 成像以及成像后数据分析。
Zotero 插件