Hello All,
I have the pellet that left over from lysing cell with Triton X-100 lysis buffer. Because I didn't sonicate the lysate and Triton X-100 lysis buffer cannot solubilize nuclear and mitochondria membranes, the pellet therefore contains cell debris, mitochondria and nucleus. I would like to isolate mitochondria, before isolating nucleus, or vice versa from this pellet to get mitochondrial protein and nuclear protein separately. Do you know any protocol to do that?
In addition, the cell that I lysed is monolayer cell line, not tissue.
Thank you so much
Cheerios,
It will not be possible as Triton X- disolves mitochondria. This paper is a good starting point: http://www.biomedcentral.com/1756-0500/5/513
Look for the following article: A simple protocol for the subcellular fractionation
of skeletal muscle cells and tissue
Also, you have to use different buffers and I dont think you can isolate it when you have used Triton-x already
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