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Updated: May 6, 2026
An Optimized Protocol to Analyze Glycolysis and Mitochondrial Respiration in Lymphocytes
Published on: November 21, 2016
Protocol for measuring cellular energetics through noncanonical amino acid tagging in human peripheral blood and
Frank Vrieling1, Hendrik J P van der Zande2, Manon Dumont3
1Division of Human Nutrition and Health, Wageningen University, 6708 WE Wageningen, the Netherlands; Department of Internal Medicine, Radboud University Medical Center, 6525 GA Nijmegen, the Netherlands.
Abstract:
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.
