cytosolic and nucleus fractionation protocol plz - (Nov/27/2007 )

hi ya..please help me to get a good protocol to fractionate the cytoplasmic and nuclic extracts....i want eliminate mumbrate proteins...,membrane proteins interuping the my area of interest protiens..thanks...

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If you want to fractionate nucleus, try centrifugation at 800-1000x g, the pellet is nucleus. The supernatant is cytoplasmic. But of course, there will be mitochondria, ER and golgi and endosomes. you have to centrifuge at 100,000x g to get the plasma membrane fraction.

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Not really, Plasma membrane comes down from 13-17Kg. It is the endosomes and such that need high centrifuge force. Of course to purify PM from the 13Kg pellet you may want to do a bit more work.

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