Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA; Feinberg Cardiovascular Research Institute, Northwestern University, Chicago, IL 60611, USA.
Department of Medicine, University of Illinois at Chicago, Chicago, IL 60612, USA.
STAR Protoc. 2022 Dec 16;3(4):101668. doi: 10.1016/j.xpro.2022.101668. Epub 2022 Sep 13.
Peritoneal macrophages (PMs) have been shown to have higher stability compared to other macrophage subtypes. However, obtaining enough PMs from a single mouse is often a limitation for metabolomics analysis. Here, we describe a protocol to isolate metabolites from a small number of mouse primary PMs for 13C-stable glucose tracing and metabolomics. Our protocol uses X for metabolite extraction instead of methanol. Our protocol can consistently extract metabolites from low cell number samples with fewer steps than methanol-based approaches. For complete details on the use and execution of this protocol, please refer to De Jesus et al., (2022).
腹腔巨噬细胞（PMs）与其他巨噬细胞亚型相比具有更高的稳定性。然而，从单个小鼠中获得足够的 PMs 通常是代谢组学分析的一个限制。在这里，我们描述了一种从少量小鼠原代 PMs 中分离代谢物的方案，用于 13C-稳定葡萄糖示踪和代谢组学分析。我们的方案使用 X 代替甲醇进行代谢物提取。与基于甲醇的方法相比，我们的方案使用更少的步骤，能够从低细胞数量的样品中稳定地提取代谢物。有关该方案使用和执行的完整详细信息，请参阅 De Jesus 等人，（2022 年）。
