Hi,
I currently using flow cytometer to performed multiple assays including Annexin V/PI staining, MitoSOX/BODIPY staining, as well as usual protein count analysis (with specific primary followed by secondary antibody). As what I know is that usually people will harvest the old media (cell treatment media) together with the trypsinized adherent cells for Ann V/PI staining assay, but how about others? As I tried to performed with either with or without and both give me different results, and I am not sure which one to trust. Thank you for your help.
Hi!
You can collect both media (detached/dead cells) and adherent cells for the analysis, however you should always use the cell death marker in your assay. Generally speaking, the dead cells will give abnormal results, as they are 'sticky' and have higher level of autofluorescence.
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