Hi,

I'm trying to obtain mitoROS levels in AML cell lines using mitoSOX in FACS. Now there are a bunch of protocols floating around on the internet and I'm trying to find the essence to what works best. My main question is regarding the staining conditions, some dilute their mitoSOX solution in PBS, some in media, some in media w/o glucose, mediua w/o FCS, PBS w/ FCS and so on. Up to this day I used to stain in PBS w/ 1 µM mitSOX for 20 minutes at 37 °C, but I worry that the glucose deprivation that comes along with this is enough to influence the mitoROS levels.

Any thoughts on this or examples of proven FACS protocols for mitoSOX?

Best,