Purification of Nucleoli From Lymphoma Cells and Solubilization of Nucleolar Proteins for 2-DE Separation

  • Régis Dieckmann
  • Yohann Couté
  • Denis Hochstrasser
  • Jean-Jacques Diaz
  • Jean-Charles Sanchez

Abstract

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).

Keywords

Saccharose Urea Lymphoma Agarose Electrophoresis 

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Copyright information

© Humana Press Inc., Totowa, NJ 2005

Authors and Affiliations

  • Régis Dieckmann
    • 1
  • Yohann Couté
    • 2
  • Denis Hochstrasser
    • 1
  • Jean-Jacques Diaz
    • 2
  • Jean-Charles Sanchez
    • 1
  1. 1.Biomedical Proteomics Research Group, Central Clinical Chemistry LaboratoryGeneva University HospitalGenevaSwitzerland
  2. 2.Centre de Genetique Moleculaire et CellulaireFrance

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