title: Evaluation and Comparison of Four Protein Extraction Protocols for Mono- and Two-Dimensional Electrophoresis inMytilus Galloprovincialis.
abstract: In this study, four protein extraction protocols fromMytilus galloprovincialiswere evaluated with the aim to identify the most practical, efficient and reproducible method. Four extraction protocols frequently used for mussels and organic matrices were selected and compared. The methods were based on the use of: i) TRIzol reagent; ii) Lysis buffer; iii) phenylmethanesulfonyl fluoride; iv) trichloroacetic acid-acetone. Protein concentration was measured by the Bradford method. Three specimens of mussels were studied and the analysis was conducted in triplicate for each of the four protocols. Results indicated that the four methods could extract significantly different protein profiles. The highest number of protein spots resolved in 2DE gels and the best reproducibility was obtained using trichloroacetic acid-acetone protocol. Results afforded the selection of a suitable extraction protocol to be used for ecotoxicoproteomics studies from mussels and for other proteomic studies conducted by particularly complex tissues such asMytilus galloprovincialis.
authors: Department of Veterinary Medicine and Animal Production, University of Naples Federico II , Italy.
authors: Marina
authors: M
authors: Ceruso
authors: Department of Veterinary Medicine and Animal Production, University of Naples Federico II , Italy.
authors: Claudia
authors: C
authors: Chirollo
authors: Department of Veterinary Medicine and Animal Production, University of Naples Federico II , Italy.
authors: Federica
authors: F
authors: Boccia
authors: Department of Veterinary Medicine and Animal Production, University of Naples Federico II , Italy.
authors: Giorgio
authors: G
authors: Smaldone
authors: Department of Veterinary Medicine and Animal Production, University of Naples Federico II , Italy.
authors: Raffaele
authors: R
authors: Marrone
authors: Department of Veterinary Medicine and Animal Production, University of Naples Federico II , Italy.
authors: Tiziana
authors: T
authors: Pepe
keywords: Mytilus galloprovincialis; Protein extraction; Proteomics