Methods to Study Phase-Separated Condensates and the Underlying Molecular Interactions.

Methods to Study Phase-Separated Condensates and the Underlying Molecular Interactions.
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DOI:
10.1016/j.tibs.2020.05.011
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发表时间:
2020-11
影响因子:
13.8
通讯作者:
Myong S
Myong S
中科院分区:
生物学1区
文献类型:
--
作者:
Ganser LR;Myong S

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已经开发了许多方法来检查通过液-液相分离(LLPS)形成的生物分子凝聚物的本体性质和潜在的分子相互作用。使用明场成像或荧光显微镜的相分离测定随时间跟踪液滴,并且可以评估各种条件对LLPS的影响,测量液滴形成和溶解动力学,并测试差异标记分子的可分辨性。光漂白后的荧光恢复(FRAP)和微流变学通过粒子跟踪探测液滴内分子的扩散,而液滴融合试验,直角成像和表面润湿确定液滴表面张力。
Many methods have been developed to examine the bulk properties and underlying molecular interactions of biomolecular condensates that form through liquid–liquid phase separation (LLPS). Phase separation assays using bright-field imaging or fluorescence microscopy track droplets over time and can assess the effect of various conditions on LLPS, measure droplet formation and dissolution kinetics, and test the miscibility of differentially labeled molecules. Fluorescence recovery after photobleaching (FRAP) and microrheology by particle tracking probe the diffusion of molecules within droplets, while droplet fusion assays, right angle imaging, and surface wetting determine droplet surface tension.
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