Hi,I tried to follow this protocol for with PBMC from CPDA buffered...

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Chris Duchenne
Jul 1, 2021
protocol Protocol: Measurement of Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) in Culture Cells for Assessment of the Energy Metabolism
Hi,

I tried to follow this protocol for with PBMC from CPDA buffered blood proveded by colleagues the day before. It worked pretty well except for a high variability and - more annoyingly - there was no proton-leakage after R/AA treatment. I guess high variability is normal for
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Sandra Loesgen Author Answered Aug 4, 2021

Department of Chemistry, Oregon State University, USA, USA,

Hi Chris, we never used PBMCs in our experiments. To my knowledge, red blood cells do not contain nuclei or mitochondria and the seahorse would not work to assess mitochondrial function. Our protocol works well with most cancer cell lines from the NCI-60 panel that we have tested. There is an article online using PBMCs, maybe check this one out?: https://movementdisorders.onlinelibrary.wiley.com/doi/pdf/10.1002/mds.104
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