基于NMR的稳定同位素追踪癌症代谢
Penghui Lin1, Andrew N Lane2, Teresa W-M Fan1
1Center for Environmental and Systems Biochemistry, Department of Toxicology and Cancer Biology, Markey Cancer Center, University of Kentucky, Lexington, KY, USA.
Methods in molecular biology (Clifton, N.J.)
|October 1, 2024
概括
核磁共振 (NMR) 能够使用稳定同位素溶解代谢学 (SIRM) 进行详细的代谢途径分析. 本协议概述了在各种生物模型中跟踪代谢动态的工作流程.
科学领域:
- 生物化学和分子生物学
- 分析化学 分析化学
- 代谢学 代谢学 代谢学
背景情况:
- 核磁共振 (NMR) 是一种强大的工具,用于对等离子体和尿液等生物流体中的代谢物 (代谢学,代谢学) 进行分析.
- 对于稳定同位素追踪器研究来说,NMR特别有价值,以阐明各种生物模型中的代谢途径,包括细胞,动物和人类受试者.
- 核磁共振的独特能力允许利用稳定的同位素来简化复杂的光谱,并量化特定原子位置的丰富度 (同位素分布分析).
研究的目的:
- 为基于NMR的稳定同位素溶解代谢学 (SIRM) 提供一个全面的协议.
- 详细介绍一个工作流程,包括前体的管理,样品的准备,以及NMR数据的获取和减少.
- 在SIRM框架内展示多重追踪器实验的应用和分析.
主要方法:
- 专注于间接检测NMR活性稳定同位素 (C,N,P,H) 使用基于H的二维NMR实验.
- 详细的工作流程,从给出同位素丰富的前体到NMR数据处理.
- 包含多重标记器实验的协议,用于增强代谢途径审讯.
主要成果:
- 成功实施全面的SIRM协议.
- 证明了对关键稳定同位素的有效间接检测.
- 应用多重追踪实验用于高级代谢分析.
结论:
- 基于NMR的SIRM为详细的代谢途径分析提供了强大的方法.
- 概述的协议有助于在各种生理和扰乱条件下研究代谢动态.
- SIRM是一种多功能技术,适用于广泛的生物系统和实验设计.
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