Protein Extraction Protocol from Tissues Treated with Formalin-Free Fixatives

The method described in this chapter allows the extraction of proteins from tissues fixed with ethanol-based fixatives (such as FineFIX®, RCL2 and methacarn) or other commercial protein-preserving fixatives. The protocol described herein is based on the use of an extraction buffer containing nonionic detergents and on the incubation of the lysate at high temperature. This method improves the extraction of those proteins that might be structurally altered or masked by the action of the fixative and enables the application of 2D analyses, due to the use of nonionic detergents that don’t modify the protein charge.