通过人外周血和小鼠组织免疫细胞中的非规范氨基酸标记测量细胞能量学的方案。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-10-08 DOI:10.1016/j.xpro.2025.104134

Frank Vrieling, Hendrik J P van der Zande, Manon Dumont, Rinke Stienstra

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引用次数: 0

摘要

细胞代谢决定了免疫细胞的功能，但我们缺乏工具来分析低产量、复杂样品的免疫代谢功能。我们提出了一种基于流式细胞术的方案，通过非规范氨基酸标记（CENCAT）在人外周血和小鼠组织免疫细胞中测量细胞能量。我们描述了样品制备、代谢抑制、蛋白质合成分析的步骤，使用点击化学、免疫分型和代谢依赖性的计算。有关本协议使用和执行的完整细节，请参阅Vrieling等人1。

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Protocol for measuring cellular energetics through noncanonical amino acid tagging in human peripheral blood and murine tissue immune cells.

Cellular metabolism dictates immune cell function, yet we lack tools to functionally profile immunometabolism in low-yield, complex samples. We present a flow cytometry-based protocol for measuring cellular energetics through noncanonical amino acid tagging (CENCAT) in human peripheral blood and murine tissue immune cells. We describe steps for sample preparation, metabolic inhibition, protein synthesis analysis using click chemistry, immunophenotyping, and calculation of metabolic dependencies. For complete details on the use and execution of this protocol, please refer to Vrieling et al.¹.

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