Proteomics

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Mass Spectrometry-compatible Subcellular Fractionation

ABSTRACT: We developed a mass spectrometry-compatible subcellular fractionation protocol. This method involves separating cells into cytoplasmic, organellar, and nuclear fractions without ultracentrifugation.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Yasushi Ishihama 

PROVIDER: PXD013581 | JPOST Repository | Fri Nov 15 00:00:00 GMT 2019

REPOSITORIES: jPOST

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Mass Spectrometry-Compatible Subcellular Fractionation for Proteomics.

Masuda Takeshi T   Sugiyama Naoyuki N   Tomita Masaru M   Ohtsuki Sumio S   Ishihama Yasushi Y  

Journal of proteome research 20191017 1

We found that nuclear envelopes stabilize against surfactants in the presence of ethylene glycol (EG). We, therefore, developed a novel subcellular fractionation approach for proteomics using RIPA buffer containing EG and phase transfer surfactants. This method involves separating the cells into the cytoplasm, organelles, and nucleus, including intermediate filaments without ultracentrifugation. These fractions are directly applicable to sample preparation for shotgun proteomics as they have no  ...[more]

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