Evaluation and comparison of four protein extraction protocols for mono- and two-dimensional electrophoresis in Mytilus galloprovincialis

  • Marina Ceruso | marina.ceruso@gmail.com Department of Veterinary Medicine and Animal Production, University of Naples Federico II, Italy.
  • Claudia Chirollo Department of Veterinary Medicine and Animal Production, University of Naples Federico II, Italy.
  • Federica Boccia Department of Veterinary Medicine and Animal Production, University of Naples Federico II, Italy.
  • Giorgio Smaldone Department of Veterinary Medicine and Animal Production, University of Naples Federico II, Italy.
  • Raffaele Marrone Department of Veterinary Medicine and Animal Production, University of Naples Federico II, Italy.
  • Tiziana Pepe Department of Veterinary Medicine and Animal Production, University of Naples Federico II, Italy.

Abstract

In this study, four protein extraction protocols from Mytilus galloprovincialis were 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 as Mytilus galloprovincialis.

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Published
2015-09-17
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Original Articles
Keywords:
Proteomics, Protein extraction, Mytilus galloprovincialis
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How to Cite
Ceruso, M., Chirollo, C., Boccia, F., Smaldone, G., Marrone, R., & Pepe, T. (2015). Evaluation and comparison of four protein extraction protocols for mono- and two-dimensional electrophoresis in Mytilus galloprovincialis. Italian Journal of Food Safety, 4(3). https://doi.org/10.4081/ijfs.2015.4534

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