Being able to determine when and under what conditions cells perform glycolysis is critical to understanding important cellular processes such as immune cell activation and cancer metabolism. However, the potential for non-glycolytic acidification can confound the interpretation of extracellular acidification data.In mammalian cell culture, the largest contributor to non-glycolytic acidification is CO2 produced by the TCA cycle.
Here we describe an approach to account for this CO2 using the mitochondrial oxygen consumption rate (measured simultaneously in the XF analyzer). Coupled with improvements to the assay design and system model, the accuracy of glycolytic rate measurement is significantly improved compared to total acidification measurements. The resulting assay reports proton efflux results directly comparable to cellular lactate efflux.
For Research Use Only. Not for use in diagnostic procedures.