Purification of Nucleoli From Lymphoma Cells and Solubilization of Nucleolar Proteins for 2-DE Separation
Differential screening of entire cell proteomes by two-dimensional gel electrophoresis (2-DE) often leads to the identification of several differentially expressed but functionally unrelated target proteins. Hence, this approach often fails to give a comprehensive picture of the underlying biological mechanism. Thus, the level of complexity of the entire cell proteome needs to be reduced (1). Cell fractionation is both a way of reducing complexity and a way to target a specific cellular compartment. The purification of nucleoli from three Hodgkin nonadherent lymphoma cell lines is presented here. It is based on an original method from Muramatsu and Onishi (2) and has been adapted from a protocol developed for HeLa cells (3).
KeywordsNucleolar Protein Dounce Homogenizer Nuclear Pellet Sucrose Cushion Intact Nucleus
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