基于点击化学方法检测原代小鼠巨噬细胞中的 4-辛基-伊他康酸烷基化蛋白质。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2024-12-20 Epub Date: 2024-09-19 DOI:10.1016/j.xpro.2024.103314

Chaofei Su, Tian Cheng, Hanyi Zhang, Hang Yin

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引用次数: 0

摘要

伊塔康酸 4-辛酯（4-OI）是伊塔康酸的一种衍生物，它通过烷基化目标蛋白质来抑制炎症。在此，我们介绍一种基于点击化学的方案，利用伊它康酸烷基（ITalk）探针检测小鼠原代骨髓源性巨噬细胞（BMDMs）中的 4-OI 烷基化蛋白质。我们介绍了培养和处理 BMDM 的步骤，以及在细胞裂解液中使用点击化学的细节。我们还详细介绍了通过 Western 印迹检测烷基化蛋白质的步骤。有关本方案使用和执行的完整细节，请参阅 Su 等人的文章1。

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Click-chemistry-based protocol for detecting 4-octyl-itaconate-alkylated proteins in primary mouse macrophages.

4-Octyl itaconate (4-OI), a derivative of itaconate, inhibits inflammation by alkylating its target proteins. Here, we present a click-chemistry-based protocol for detecting 4-OI-alkylated proteins in mouse primary bone-marrow-derived macrophages (BMDMs) by using an itaconate-alkyne (ITalk) probe. We describe steps for culturing and treating BMDMs and details on using click chemistry in the cell lysate. We also detail procedures for detecting alkylated proteins by western blot. For complete details on the use and execution of this protocol, please refer to Su et al.¹.

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