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Sample sonication after trichloroacetic acid precipitation increases protein recovery from cultured hippocampal neurons, and improves resolution and reproducibility in two-dimensional gel electrophoresis

Authors :
Carlos B. Duarte
Bruno Manadas
Konstantinos Vougas
Michael Fountoulakis
Source :
Repositório Científico de Acesso Aberto de Portugal, Repositório Científico de Acesso Aberto de Portugal (RCAAP), instacron:RCAAP
Publication Year :
2006

Abstract

Protein precipitation with TCA followed by acetone washing is frequently used to clean samples before 2-DE. However, the difficulty in solubilizing TCA-precipitated proteins causes some variability in 2-D gels and makes it difficult to detect some proteins. In this work we show that sonication of the samples, after TCA precipitation followed by elution in sample buffer, increases total protein recovery, and improves reproducibility and matching ratios between gels when analyzed by specialized software.

Details

ISSN :
01730835
Volume :
27
Issue :
9
Database :
OpenAIRE
Journal :
Electrophoresis
Accession number :
edsair.doi.dedup.....5130a6771cedfe765058057e885585e1