Subcellular fractionation

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verajyl
verajyl's picture
Subcellular fractionation

Hi,

I am looking into methods to isolate mitochondria and nuclei from cultured cells. I have already isolated plasma membrane from the post-nuclear supernatant, and still have the nuclear fractions (after removing the post-nuclear fraction). I would like to isolate and separate mitochondria and nuclei from whatever that is left. Is there a protocol to do so? Looking forward to your comments and advice!

Thanks!

heehawmcduff
heehawmcduff's picture
Hi there,

Hi there,

I have used this kit with some degree of success.  The isolation procedure is pretty easy and the mitochondria were intact and could be manipulated further.  Western blots confirmed that it yields pure fractions too.

http://www.piercenet.com/products/browse.cfm?fldID=B32ABB4E-5056-8A76-4EF4-60A04BB784E7

Best wishes

Biju
Biju's picture
Hi

Hi
I hav tried ultracentrifugation to separate mitochondrial fraction from peripheral blood and did activity staining using the isolated fraction.
Biju Joseph