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Comparison of protein precipitation methods for various rat brain structures prior to proteomic analysis
Author(s) -
Fic Ewelina,
KedrackaKrok Sylwia,
Jankowska Urszula,
Pirog Artur,
DziedzickaWasylewska Marta
Publication year - 2010
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201000197
Subject(s) - acetone , protein precipitation , precipitation , chloroform , sonication , chromatography , chemistry , proteomics , methanol , protein purification , sample preparation , biochemistry , mass spectrometry , organic chemistry , physics , meteorology , gene
Sample preparation is a fundamental step in proteomic methodologies. The quality of the results from a proteomic experiment is dependent on the nature of the sample and the properties of the proteins. In this study, various pre‐treatment methods were compared by proteomic analysis; we analysed various rat brain structures after chloroform/methanol, acetone, TCA/acetone and TCA protein precipitation procedures. The protein content of the supernatant was also examined by 2‐DE. We found that for four of the rat brain structures, precipitation with chloroform/methanol and acetone delivered the highest protein recovery for top‐down proteomic analysis; however, TCA precipitation resulted in good protein separation and the highest number of protein spots in 2‐DE. Moreover, TCA precipitation also gave high efficiency of protein recovery if prior sonication procedure was performed.