I have some questions regarding to buffer and method for cultured cell fractionation to get mitochondrial, nuclear, cytosol proteins as following:
- Can I just use PBS with some inhibitors for buffer? because what matter is centrifuge speed to dissociate cell organelles.
- Can I use just mild lysis buffer (NP40 or tritonx100) to homoginize cell lysate instead of douce homoginizer before centrifuge each pellet?
- There are so many recipe for the buffer and method for cell fractanation, which is the best or most standard protocol I should follow? Any paper suggestion?
Thank you so much
Edited by asaren, 09 May 2015 - 08:11 PM.