Measuring NLR Oligomerization III: Detection of NLRP3 Complex by Bioluminescence Resonance Energy Transfer
Measuring NLR Oligomerization III: Detection of NLRP3 Complex by Bioluminescence Resonance Energy Transfer
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DOI：
10.1007/978-1-4939-3566-6_10
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发表时间：
2016-01-01
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影响因子：
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通讯作者：
Pelegrin, Pablo
中科院分区：
文献类型：
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作者：
Martin-Sanchez, Fatima;Compan, Vincent;Pelegrin, Pablo
生物发光共振能量转移(BRET)是生物发光供体(Renilla荧光素酶)和荧光蛋白受体之间非辐射能量转移的一种自然现象。BRET信号依赖于供体和受体之间的距离和方向，可以用来实时研究活细胞中蛋白质之间的相互作用和蛋白质内部的构象变化。本方案描述了使用BRET技术来研究活细胞中NLRP3在炎症小体激活之前和期间的寡聚。
Bioluminescent resonance energy transfer (BRET) is a natural phenomenon resulting from a non-radiative energy transfer between a bioluminescent donor (Renilla luciferase) and a fluorescent protein acceptor. BRET signal is dependent on the distance and the orientation between the donor and the acceptor and could be used to study protein-protein interactions and conformational changes within proteins in real time in living cells. This protocol describes the use of BRET technique to study NLRP3 oligomerization in living cells before and during NLRP3 inflammasome activation.