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.
Thanks.
-Blotman-
QUOTE (Blotman @ Feb 13 2008, 12:44 PM)
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.
Thanks.
Thanks.
lose-fit pottering and differential centrifugation
-The Bearer-
QUOTE (The Bearer @ Feb 13 2008, 02:04 PM)
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.
-Blotman-