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Two-dimensional gel electrophoresis of peripheral nerve proteins: Optimized sample preparation

Authors :
Maria Le Pera
Elena Urso
Paola Valentino
Antonio Qualtieri
Massimo Scornaienchi
Francesco Bono
Aldo Quattrone
Source :
Journal of neuroscience methods 159 (2007): 125–133. doi:10.1016/j.jneumeth.2006.07.00, info:cnr-pdr/source/autori:A.Qualtieri, M. Le Pera, E.Urso, F. Bono, P. Valentino, MC. Scormaienchi, A.Quattrone/titolo:Two-dimensional gel electrophoresis of peripheral nerve proteins: Optimized sample preparation./doi:10.1016%2Fj.jneumeth.2006.07.00/rivista:Journal of neuroscience methods/anno:2007/pagina_da:125/pagina_a:133/intervallo_pagine:125–133/volume:159
Publication Year :
2006

Abstract

For proteomic analysis, sample preparation plays a crucial role in two-dimensional gel electrophoresis (2DE), since, very often, each tissue or cell culture requires specific treatments. In the present paper, we report a sample preparation procedure suitable for 2DE that was done on peripheral nerve using bovine sciatic nerves and human sural nerve biopsies. We obtained an appreciable reduction of tissue heterogeneity using protein extracts obtained from nerve-fiber bundles instead of the entire nerve. In addition, we optimized 2DE protein separation using a combination of CHAPS, Triton X-100, and SB3-10 detergents in an isoelectric-focusing (IEF) buffer. The reported experimental procedures appear to be essential for 2DE separation of peripheral nerve proteins for the establishment of a reference map.

Details

ISSN :
01650270
Volume :
159
Issue :
1
Database :
OpenAIRE
Journal :
Journal of neuroscience methods
Accession number :
edsair.doi.dedup.....f7c3c64a17cfac249167ef5f24466458
Full Text :
https://doi.org/10.1016/j.jneumeth.2006.07.00