Chapter title |
Full-Length Protein Extraction Protocols and Gel-Based Downstream Applications in Formalin-Fixed Tissue Proteomics
|
---|---|
Chapter number | 11 |
Book title |
Proteomic Profiling
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2550-6_11 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2549-0, 978-1-4939-2550-6
|
Authors |
Alessandro Tanca, Sergio Uzzau, Maria Filippa Addis |
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. |
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