Fluorescence polarization and time-resolved fluorescence resonance energy transfer techniques for PI3K assays.
Fluorescence polarization and time-resolved fluorescence resonance energy transfer techniques for PI3K assays.
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用于 PI3K 测定的荧光偏振和时间分辨荧光共振能量转移技术。
DOI：
10.1007/978-1-60761-244-5_10
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Ma,Haiching
中科院分区：
文献类型：
--
作者：
Horiuchi,KurumiY;Ma,Haiching
基于荧光的生化检测灵敏且使用方便；因此，它们广泛用于酶测定和分子相互作用研究。然而，当将该方法应用于筛选用于药物发现的化合物库时，化学库中通常大量存在的高荧光化合物是有问题的。荧光偏振 (FP) 和时间分辨荧光共振能量转移 (TR-FRET) 检测受化合物荧光影响较小，适合大规模高通量筛选 (HTS)。在本节中，我们描述 PI3 激酶 (PI3K) 的同质 FP 和 TR-FRET 方法，PI3K 是一种“难以进行 HTS”的脂质激酶家族，因为传统的放射性同位素测定很难应用于 HTS 格式。将描述 FP 和 TR-FRET 技术在 PI3K HTS 中的应用，并讨论这些测定的优点和缺点。
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.