Abstract
Staining of two-dimensional gels is a primary concern in proteomic studies using two-dimensional gel electrophoresis with respect to the number of proteins analyzed, the accuracy of spot quantification and reproducibility. In this review article, the efficiency of the most widely used dyes was investigated. Visible dyes (Coomassie blue and silver nitrate), fluorescent dyes (Sypro Ruby, Deep Purple) and cyanine labeled methods were compared.
Highlights
Protein separation by two-dimensional electrophoresis (2DE) is largely used in proteomic approaches because of both high resolution and the availability of powerful image analysis software for gel comparison and compatibility with subsequent protein characterization by mass spectrometry [1]
The present review compares several non-covalent and covalent protein dyes for large-scale comparison of 2D gel electrophoresis patterns by image analysis in order to select the best method according to the biological question and sample type
The binding behavior is attributed to Van der Waals forces and hydrophobic interactions. Such non-covalent binding of the dye allowed an excellent compatibility with MALDI-TOF mass spectrometry [12]
Summary
Protein separation by two-dimensional electrophoresis (2DE) is largely used in proteomic approaches because of both high resolution and the availability of powerful image analysis software for gel comparison and compatibility with subsequent protein characterization by mass spectrometry [1]. For these various aspects, the selection of the protein staining procedure is of major importance [2]. Covalent protein labeling, called ‘‘difference gel electrophoresis’’ (DIGE) is nowadays an interesting alternative, which allowes multiplexing of samples and the use of an internal standard [10] This pre-electrophoretic protein stain method leads to highly accurate qualitative and quantitative results, because gel-to-gel variations are eliminated. The present review compares several non-covalent and covalent protein dyes for large-scale comparison of 2D gel electrophoresis patterns by image analysis in order to select the best method according to the biological question and sample type
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