Köster Mario, Frahm Thomas, Hauser Hansjörg
Department of Gene Regulation and Differentiation, GBF, Mascheroder Weg 1, D-38124 Braunschweig, Germany.
Curr Opin Biotechnol. 2005 Feb;16(1):28-34. doi: 10.1016/j.copbio.2004.11.002.
Protein mobility within cells is of key importance for many cellular functions. Although immunostaining can reveal protein locations in the steady-state, this might not represent the full picture and provides no information about protein movements. Fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP) are two techniques that enable the dynamics of intracellular protein mobility to be studied. These technologies have been successfully used to analyze the nucleocytoplasmic shuttling of STAT1, an intracellular signal transducer and activator of transcription, and can applied to the study of other proteins. Furthermore, FRAP and FLIP approaches have the added advantage of not affecting cell viability and might find application in the imaging of intracellular events in certain tissues and live animals.
蛋白质在细胞内的移动性对许多细胞功能至关重要。尽管免疫染色可以揭示蛋白质在稳态下的位置，但这可能并不代表全貌，也无法提供有关蛋白质运动的信息。光漂白后荧光恢复（FRAP）和光漂白荧光损失（FLIP）是两种能够研究细胞内蛋白质移动性动态的技术。这些技术已成功用于分析细胞内信号转导子和转录激活子STAT1的核质穿梭，也可应用于其他蛋白质的研究。此外，FRAP和FLIP方法还有不影响细胞活力的额外优势，可能在某些组织和活体动物的细胞内事件成像中得到应用。
