Archival formalin-fixed, paraffin-embedded (FFPE) tissue repositories and their associated clinical information can represent a valuable resource for tissue proteomics. In order to make these tissues available for protein biomarker discovery and validation studies, dedicated sample preparation procedures overcoming the intermolecular cross-links introduced by formalin need to be implemented. This chapter describes a full-length protein extraction protocol optimized for downstream gel-based proteomics applications. Using the procedures detailed here, SDS-PAGE, western immunoblotting, GeLC-MS/MS, 2D-PAGE, and 2D-DIGE can be carried out on FFPE tissues. Technical tips, critical aspects, and drawbacks of the method are presented and discussed.

Full-length protein extraction protocols and gel-based downstream applications in formalin-fixed tissue proteomics / A. Tanca, S. Uzzau, M.F. Addis (METHODS IN MOLECULAR BIOLOGY). - In: Proteomic Profiling : Methods and Protocols / [a cura di] A. Posch. - [s.l] : Humana Press, 2015. - ISBN 9781493925490. - pp. 117-134

Full-length protein extraction protocols and gel-based downstream applications in formalin-fixed tissue proteomics

M.F. Addis
Ultimo
2015

Abstract

Archival formalin-fixed, paraffin-embedded (FFPE) tissue repositories and their associated clinical information can represent a valuable resource for tissue proteomics. In order to make these tissues available for protein biomarker discovery and validation studies, dedicated sample preparation procedures overcoming the intermolecular cross-links introduced by formalin need to be implemented. This chapter describes a full-length protein extraction protocol optimized for downstream gel-based proteomics applications. Using the procedures detailed here, SDS-PAGE, western immunoblotting, GeLC-MS/MS, 2D-PAGE, and 2D-DIGE can be carried out on FFPE tissues. Technical tips, critical aspects, and drawbacks of the method are presented and discussed.
2d-page; archival tissues; biobanks; dige; electrophoresis; ffpe; gelc-ms/ms; immunoblotting; mass spectrometry; paraffin-embedded; proteomics; blotting, western; chromatography, liquid; electrophoresis, gel, two-dimensional; proteins; proteomics; tandem mass spectrometry; two-dimensional difference gel electrophoresis; molecular biology; genetics
Settore BIO/10 - Biochimica
2015
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/492729
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