Glycolytic rate assay with PBMCs

Hello, we are setting up a seahorse assay for a immunology master class. We want to measure the glycolytic rate in human PBMCs or purified human CD4 T cells activated vs non activated. We fail to see any difference. Has anyone done this kind of assay and saw differences in the glycolytic rate? Would be willing to share a protocol? We have tried after overnight stimulation with soluble CD3/CD28 and also injecting the stimuli in the port A and follow the ECAR for 1h before the injection of the inhibitors. Our results with T cells are particularly striking as it seems they are completely quiescent and the addition of the 2-DG seems to even increase ECAR on some wells.

Thanks for any help. As I said this is for a master course in immunology and the students are quite disappointed when we never see effects ...

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