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Two-dimensional gel electrophoresis of peripheral nerve proteins: Optimized sample preparation
- 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.
- Subjects :
- Detergents
Sural nerve
Nerve Tissue Proteins
Biology
Peptide Mapping
Specimen Handling
Myelin
Nerve Fibers
Sural Nerve
Chaps
Protein purification
medicine
Animals
Humans
Sample preparation
Electrophoresis, Gel, Two-Dimensional
Peripheral Nerves
Gel electrophoresis
Chromatography
Two-dimensional gel electrophoresis
General Neuroscience
medicine.anatomical_structure
Cell culture
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Cattle
Gels
Subjects
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