title: Full-length protein extraction protocols and gel-based downstream applications in formalin-fixed tissue proteomics.
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.
authors.affiliation: Porto Conte Ricerche, Loc, S.P. 55 Porto Conte/Capo Caccia Km 8.400, Tramariglio, Alghero (SS), 07041, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Tanca
authors.forename: Sergio
authors.initials: S
authors.lastname: Uzzau
authors.forename: Maria Filippa
authors.initials: MF
authors.lastname: Addis
mesh_terms: D015153:Blotting, Western; D002853:Chromatography, Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry; D057825:Two-Dimensional Difference Gel Electrophoresis
chemical_list: D011506:Proteins