Cell Brakeage...how will I know - I need the mitochondria. (Aug/18/2009 )

Im isolating Mitochondria from HEP G2 cells and Im new to all of this. I have a kit but its a bit old.

I have cells that turn blue under after trypan blue staining but they look largely intact to me under a microscope and Im disspointed that I dont seem to have mitochondria. Can someone advise me as to what properly broken cells should look like?? Should there be fragmentation?

Best regards,

the john

You will need to disrupt the cells to extract the mitochondria. The blue, but intact looking cells you are seeing are not disrupted enough. You don't need kits to get mitos, there are plenty of freely available protocols for sub-cellular fractionation.