1 reply to this topic

[sowmya]

Hi everybody,

I have been trying to isolate mitochondria and mitochondria associated membranes from T cells. I used mitochondria isolation kit for isolation of mitochondria wherein i see a pellet. I dissolved this pellet in resupension buffer and when i did ultracentrifugation using amersham percoll at 95000 g for 30 min, I dont see any band for mitochondria and mito associated membranes. To start with I take 10^8 activated T-cells. I would also like to know if there is lifespan for percoll? I am using percoll which had got expired a year ago! will it affect the process of seperation?

Thanks in advance

[Inmost sun]

sowmya, on Apr 10 2010, 12:18 PM, said:

Hi everybody,

I have been trying to isolate mitochondria and mitochondria associated membranes from T cells. I used mitochondria isolation kit for isolation of mitochondria wherein i see a pellet. I dissolved this pellet in resupension buffer and when i did ultracentrifugation using amersham percoll at 95000 g for 30 min, I dont see any band for mitochondria and mito associated membranes. To start with I take 10^8 activated T-cells. I would also like to know if there is lifespan for percoll? I am using percoll which had got expired a year ago! will it affect the process of seperation?

Thanks in advance

Did you perform density gradient centrifugation?

The question for me is if the percoll was open or sealed. The water content may have changed.