费斯特共振能量转移
荧光
荧光各向异性
共振荧光
荧光互相关光谱
能量转移
极化(电化学)
生命科学中的荧光
核磁共振
材料科学
化学
荧光光谱法
物理
光学
分子物理学
物理化学
作者
Kazuo Horiuchi,Haiching Ma
出处
期刊:Methods in molecular biology
日期:2009-10-03
卷期号:: 161-176
被引量:1
标识
DOI:10.1007/978-1-60761-244-5_10
摘要
Fluorescence-based biochemical assays are sensitive and convenient to use; therefore, they are widely employed for enzyme assays and molecular interaction studies. However, when this method is applied for screening of a compound library for drug discovery, high fluorescence compounds, which usually exist in large numbers in chemical libraries, are problematic. Fluorescence Polarization (FP) and Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) assays are less affected by compound fluorescence and suitable for large-scale high-throughput screening (HTS). In this section, we describe homogenous FP and TR-FRET methods for PI3-kinase (PI3K), a family of lipid kinases that is “difficult-to-do-HTS” since traditional radioisotope assays are hard to apply to HTS format. The application of FP and TR-FRET techniques for PI3K HTS will be described and advantages and disadvantages of these assays will be discussed.
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