荧光相关光谱
荧光
化学
荧光互相关光谱
过氧化物酶
过氧化氢
葡萄糖氧化酶
荧光光谱法
生命科学中的荧光
光谱学
分析化学(期刊)
生物物理学
分子
光化学
生物化学
色谱法
生物传感器
酶
生物
有机化学
光学
物理
量子力学
作者
Etsuro Ito,Shigeo Watabe,Mika Morikawa,Hiromi Kodama,Ryuichi Okada,Toshiaki Miura
出处
期刊:Methods in Enzymology
日期:2013-01-01
卷期号:: 135-143
被引量:14
标识
DOI:10.1016/b978-0-12-405883-5.00008-9
摘要
Fluorescence correlation spectroscopy (FCS) is a technique in which measurement of fluorescence intensity fluctuations is used to clarify dynamic molecular interactions within a very small space in a solution containing a small number of fluorescent molecules. The FCS-based analysis gives the average number and average diffusion time of the fluorescent molecules during their passage through a very small space. One advantage of FCS is that physical separation between free and bound fluorescent probes is not required because the properties of fluorescence fluctuations are accounted for. Therefore, when fluorescent probes are bound with proteins by peroxidase and hydrogen peroxide (H2O2), FCS enables us to detect H2O2 with high sensitivity. In addition, because H2O2 is generated by oxidase-catalyzed reactions, a highly sensitive method for detecting H2O2 is applicable to the measurement of low levels of various oxidases and their substrates, such as glucose. We here describe the protocol of a de novo, highly sensitive method for the measurement of H2O2 and glucose using FCS.
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