Optimized protocol to isolate primary mouse peritoneal macrophage metabolites

Autor: Adam De Jesus, Carolina M. Pusec, Tivoli Nguyen, Farnaz Keyhani-Nejad, Peng Gao, Samuel E. Weinberg, Hossein Ardehali
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: STAR Protocols, Vol 3, Iss 4, Pp 101668- (2022)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2022.101668
Popis: Summary: 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). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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