Isolation of a clean cytoplasmic fraction - (Feb/13/2008 )
I am trying to fractionate mammalian cells to obtain cytoplasmic, soluble nuclear, chromatin and nuclear matrix fractions. My problem is that the nuclear fraction is leaking into the cytoplasmic fraction. Is there a good recipe to obtain a clean cytoplasmic fraction. I would not mind even if I lost the other fractions.
lose-fit pottering and differential centrifugation
Could you please explain this in more detail? What is loose-fit pottering?? And how will differential centrifugation help.. and at what speeds? Could you please point me to a protocol?
Thank you very much.