Standard Dyes for Total Protein Staining in Gel-Based Proteomic Analysis 英文参考文献.docVIP

Materials 2010, 3, 4784-4792; doi:10.3390/ma3104784 OPEN ACCESS materials ISSN 1996-1944 /journal/materials Review Standard Dyes for Total Protein Staining in Gel-Based Proteomic Analysis Fran?ois Chevalier Proteomic Laboratory, iRCM, CEA, Fontenay aux Roses, France; E-Mail: francois.chevalier@cea.fr; Tel.: +33-146-548-326; Fax: +33-146-549-138. Received: 8 October 2010; in revised form: 13 October 2010 / Accepted: 15 October 2010 / Published: 20 October 2010 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. Keywords: protein staining; fluorescent dyes; two-dimensional electrophoresis; proteomics 1. Introduction 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]. Based on two independent biochemical characteristics of proteins, 2DE combines isoelectric focusing, which separates proteins according to their isoelectric point, and SDS, which separates them further according to their molecular mass (Figure 1, step 2). The next typical steps of the flow of gel-based proteomics are spots visualization and evaluation (Figure 1, step 3), expression analysis, and finally protein identification by mass spectromet