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Purpose: Recent developments in MS technology enable comprehensive proteome analysis of relatively small tissue samples of human cancer or tissue- or stage-specific differential expression in humans as well as analysis of biogenic molecules in metabolic pathways. However, the detailed subcellular distribution of the proteins in a cell remains unknown. Objective: We herein report a simple subcellular fractionation method that has been optimized using colorectal cancer cell line models. Methodology: We developed a membrane permeabilization method to isolate cellular organelles, and then applied this method to fractionate three colorectal cancer cell lines to systematically investigate the distribution of the proteins. Results: We isolated membrane, cytoplasm, and cytoskeletons fraction from three colorectal cancer cell lines. Proteins in membrane and cytoskeleton fractions were less abundant than those in cytosolic fractions. Immunohistochemistry showed that proteins localized to a particular organelle, regardless of their fractionation. Conclusions: Here, we identified 506 cellular organelle proteins by using our subcellular protein fractionation method. This fractionated protein list was useful for investigating the subcellular localization of any protein on a global scale. Highlights: We identified 506 cellular organelle proteins by using our subcellular protein fractionation method. This fractionated protein list was useful for investigating the subcellular localization of any protein on a global scale.With the tools of modern technology like filming and editing video, it is now more accessible than ever to combine your subjects’ voices, images, graphics, texts, and video into a unified presentation. This course will introduce
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