I have tried several protocols for mitochondria extraction but none of these were adapted for correctly visualizing by western-blot, translocation of Drp1 protein from cytosolic fraction to mitochondrial fraction.
In other words, has anyone an adapted mitochondrial extraction protocol adapted for this purpose ? Thank's!!!
Hello Daniel,
please check Fig. 3 c, d in the attached paper. The authors investigated the translocation of DRP1 from the cytosol to mitochondria. In the supplementary file you can find information how they performed the analysis.
I hope this helps you.
Best regards, Christian
Hello Daniel,
I have done western blotting for DRP1 (and other proteins) in the mitochondrial fraction routinely. You can find our protocol (heavy membrane isolation) in the paper below. This has worked very well for us with multiple cell lines.
Good luck!
Madhavika
http://www.ncbi.nlm.nih.gov/pubmed/24608435
Thank you Madhavika,
I think that the most critical point is the number of cells to extract.
regards,
Daniel
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