• Evaluation of two methods for protein extraction from muscle tissues. • Two different extraction methods showed distinct patterns of protein abundance; • Two-dimensional electrophoresis revealed 131 spots with different volumes; • SDS-based buffer (method A) mostly extracted proteins of high MW range and peculiar in sarcomere and muscular fibres, such as Troponin, Myosin, Miozenin 2 and Ankyrin; • Urea/chaps/DTE/tris solution (method B) enriched numerous protein species in a low MW range, several protein fragments and organelle membrane proteins