Two-dimensional gel electrophoresis of peripheral nerve proteins: Optimized sample preparation.
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.
Qualtieri
Antonio
A
Institute of Neurological Sciences, National Research Council (CNR), Contrada Burga, 87050 Mangone, Cosenza, Italy. a.qualtieri@isn.cnr.it
Le Pera
Maria
M
Urso
Elena
E
Bono
Francesco
F
Valentino
Paola
P
Scornaienchi
Massimo Corrado
MC
Quattrone
Aldo
A
D000818:Animals; D002417:Cattle; D003902:Detergents; D015180:Electrophoresis, Gel, Two-Dimensional; D005782:Gels; D006801:Humans; D009412:Nerve Fibers / Q000737:chemistry; D009419:Nerve Tissue Proteins / Q000032:analysis*; D010449:Peptide Mapping; D010525:Peripheral Nerves / Q000737:chemistry*; D013048:Specimen Handling; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013497:Sural Nerve / Q000737:chemistry
D003902:Detergents; D005782:Gels; D009419:Nerve Tissue Proteins