Hello all, I am trying to perform subcellular fractionation in onion cells. I want to separate membrane, cytosol and nuclear compartments. Has any one tried this using lab-made buffer and protocols? what precautions does one have to take in order to prevent cross-contamination ? I would be grateful if someone could share a well standardized, an established protocol and reagents for subcellular fractionation in onion cells. Also, I would like to know what markers (in onion cells) could be used to validate if the subcellular fractionation have been accurate? Please suggest.

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