Hello,
Some said that the sonication can break nuclear membrane and get the nuclear protein. However many protocol also use the sonication method to disrupt cell before isolating mitrochondrial fraction. So, I confuse if the sonication can solubilize both nuclear and mitochondrial membrane or just only soubilize nuclear membrane? If so why the sonication cannot solubilize mitochondrial membrane?
Thank you
sonication would break all kinds of membranes irrespective of mitochondrial or nuclear thus releases proteins from nucleus and mitochondria (and other organelles). If you need to isolates proteins from different organelles/cellular fractions than it would be best to use some cell-fractionation kits. Good luck.
A short sonication with low energy can disrupt the plasma membrane without breaking too many mitochondrial or nuclear membranes yielding in tact mitochondria.
I think the best way to do this is to swell the cells on ice for 15-20 minutes in a hypotonic solution and either run a few times through a 25 gauge needle or dounce homogenize.
I separate lysosomal, mitochondrial and nuclear fractions with swelling, dounce homogenization and differential centrifugation in Opti-Prep.
Hope this helps
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