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2-D PAGE and MS analysis of proteins from formalin-fixed, paraffin-embedded tissues

Addis, Maria Filippa and Tanca, Alessandro and Pagnozzi, Daniela and Rocca, Stefano and Uzzau, Sergio (2009) 2-D PAGE and MS analysis of proteins from formalin-fixed, paraffin-embedded tissues. Proteomics, Vol. 9 (18), p. 4329-4339. eISSN 1615-9861. Article.

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DOI: 10.1002/pmic.200900010

Abstract

In the past decade, encouraging results have been obtained in extraction and analysis of proteins from formalin-fixed, paraffin-embedded (FFPE) tissues. However, 2-D PAGE protein maps with satisfactory proteomic information and comparability to fresh tissues have never been described to date. In the present study, we report 2-D PAGE separation and MS identification of full-length proteins extracted from FFPE skeletal muscle tissue. The 2-D protein profiles obtained from FFPE tissues could be matched to those achieved from frozen tissues replicates. Up to 250 spots were clearly detected in 2-D maps of proteins from FFPE tissue following standard mass-compatible silver staining. Protein spots from both FFPE and frozen tissue 2-D gels were excised, subjected to in situ hydrolysis, and identified by MS analysis. Matched spots produced matched protein identifications. Moreover, 2-D protein maps from FFPE tissues were successfully subjected to Western immunoblotting, producing comparable results to fresh-frozen tissues. In conclusion, this study provides evidence that, when adequately extracted, full-length proteins from FFPE tissues might be suitable to 2-D PAGE-MS analysis, allowing differential proteomic studies on the vast existing archives of healthy and pathological-fixed tissues.

Item Type:Article
ID Code:3590
Status:Published
Refereed:Yes
Uncontrolled Keywords:Proteomics, FFPE tissue, 2D-PAGE
Subjects:Area 06 - Scienze mediche > MED/07 Microbiologia e microbiologia clinica
Area 07 - Scienze agrarie e veterinarie > VET/03 Patologia generale e anatomia patologica veterinaria
Divisions:001 Università di Sassari > 01 Dipartimenti > Patologia e clinica veterinaria
001 Università di Sassari > 01 Dipartimenti > Scienze biomediche
002 Altri enti e centri di ricerca del Nord Sardegna > Porto Conte Ricerche, Alghero
Publisher:Wiley
eISSN:1615-9861
Deposited On:03 Mar 2010 11:51

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