Convenient and versatile subcellular extraction procedure, that facilitates classical protein expression profiling and functional protein analysis |
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Authors: | Abdolzade-Bavil Afsaneh Hayes Scott Goretzki Lothar Kröger Michaela Anders Jonas Hendriks Robertus |
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Affiliation: | Merck, Life Science Products R&D, Darmstadt, Germany. |
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Abstract: | Standardized sample preparation to reduce proteome complexity facilitates subsequent proteome analysis. Here we describe a robust sequential extraction method that enables simple fractionation of proteins in their native state according to their subcellular localization, yielding four subproteomes enriched in (a) cytosolic; (b) membrane and membrane organelle-localized; (c) soluble and DNA-associated nuclear and (d) cytoskeletal proteins. Efficiency and selectivity is demonstrated by morphological-, two-dimensional electrophoresis image-, immunological- as well as enzymatic-analysis. In pilot studies, subcellular redistribution of regulatory proteins was successfully measured. |
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