Mitochondrial Isolation; Anyone done it?

[MSTPbound]

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Just wondering if anyone has (successfully) extracted mitochondria - if so, can you recommend a protocol? Actually, I am able to get them at the interphase of two buffers, but I'm having trouble figuring out the best way to get them out without pulling too much buffer (used a 27G needle, but still pulled too much buffer).

Thanks.

-MSTPbound

 

[MSTPbound]

Ok... I guess that means no. I finally figured it out; just had to break a 1-mL pipet in half and play with it a little bit.

 

[Fuzuli]

I haven't extracted mitochondria yet, but here's a couple of links from www.protocol-online.org:


http://www.protocol-online.org/prot/Cell_Biology/Cell_Parts___Organelles/Mitochondria/: The main site
http://cmm.ucsd.edu/Lab_Pages/Dixon/01/Protocols_IsolationOfRatLiverMito.html
http://www.abcam.com/assets/pdf/protocols/Mitochondrial purification protocol.pdf

Good luck.

 

[stochastic]

I didn't check these links, but I have had success purifying them with Dynal M-500 magnetic beads conjugated to anti-porin antibodies.