I've performed some Seahorse analyses on IPEC-J2 cells. However, I noticed that the basal OCR values were rather low when conducting the ATP rate assay (around 8 pmol/min). When repeating the experiment, I increased the cell density a bit (the cells were almost confluent at the time of analysis). The basal OCR values were then increased a bit (around 20 pmol/min), but still quite low. When performing a mito stress test, the basal OCR values were much higher (between 65 and 80 pmol/min). The only difference between both assays (besides the loading of the cartridges) is that the last washing step needs to be done right before the start of the ATP rate assay. Is it possible that this last washing step could cause the observed low basal OCR values?
Thank you in advance.