I have a protein which might be dual localized in the mitochondria and in the cytoplasm. Therefore, I have to get them separated and check on the Western blot in which of these fractions I see my protein. Any help?
TIA
Charles D Anderson
Dear Maya,
I attached some of my protocols i have used in the past for a different cell lines.
Some variation in fractional location observed with different example as, with swelling cells and douncing, or with 26G needle and syringe up and down diffrent number of times vs using nitrogen decavitation chamber to lyse cells. It seemed the best for me at this time was to get a clean ER fraction, which we used a chamber for nitrogen decavitation, with discontinuous sucrose gradient.
Best Wishes!
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