Mitochondria extraction - starting amounts (Apr/24/2009 )

Hi,

I'm a beginner in mitochondria extraction and I'm hoping to get some advice.
I've got the kit from pierce, which using a reagent based method for extraction (no homogeniser needed)

Normally, what's a good amount of starting material from cell lines?

I've started with a T75, fully confluent, but I didnt seem to get any mitochondria pellet?

Thanks

Sharon Pek on Apr 24 2009, 03:46 AM said:

Depends on how much mitochondia you need.

If you're using very few cells, then perhaps the pellet is too small to see. Have a good light microscope? If so, take a look.

I hate to suggest this, because I know that people buy the kits specifically to avoid doing this, but if you've tried the kit a few times (scaling up the amount of cells used) and haven't got anything perhaps you should double-check using the homogenization method. If you use two methods and you still can't get mitochondria, then it's probably not a problem with the method and you can start troubleshooting other aspects of your experiment.

Let me know how things work out.

Cheers,
-Carlton