From a 2DE-gel spot to protein function: Lesson learned from HS1 in chronic lymphocytic leukemia

Benedetta Apollonio, Maria Teresa Sabrina Bertilaccio, Umberto Restuccia, Pamela Ranghetti, Federica Barbaglio, Paolo Ghia, Federico Caligaris-Cappio, Cristina Scielzo

Research output: Contribution to journalArticlepeer-review

Abstract

The identification of molecules involved in tumor initiation and progression is fundamental for understanding disease’s biology and, as a consequence, for the clinical management of patients. In the present work we will describe an optimized proteomic approach for the identification of molecules involved in the progression of Chronic Lymphocytic Leukemia (CLL). In detail, leukemic cell lysates are resolved by 2-dimensional Electrophoresis (2DE) and visualized as “spots” on the 2DE gels. Comparative analysis of proteomic maps allows the identification of differentially expressed proteins (in terms of abundance and post-translational modifications) that are picked, isolated and identified by Mass Spectrometry (MS). The biological function of the identified candidates can be tested by different assays (i.e. migration, adhesion and F-actin polymerization), that we have optimized for primary leukemic cells.

Original languageEnglish
Article numbere51942
JournalJournal of visualized experiments : JoVE
Issue number92
DOIs
Publication statusPublished - Oct 19 2014

Keywords

  • 2D Electrophoresis
  • Chronic lymphocytic leukemia
  • Cytoskeleton
  • Issue 92
  • Lymphocytes
  • Mass spectrometry
  • Medicine
  • Migration

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)
  • Medicine(all)

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