Journal
EXPERT REVIEW OF PROTEOMICS
Volume 9, Issue 6, Pages 627-634Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1586/EPR.12.63
Keywords
2D gel electrophoresis; colloidal coomassie brilliant blue; comparative proteomics; image analysis; immobilized pH gradient; mass spectrometry; protein identification; protein staining
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Funding
- Progetti di ricerca di Ateneo from the University of Genoa [N CUP D31J11001610005]
- Fondazione Mara Wilma e Bianca Querci, la Nuova Speranza and Compagnia di San Paolo for G Candiano
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Gel-based proteomics are the most useful method for protein separation, even when compared with gel-free proteomics. Proteomic analysis by 2D gel electrophoresis (2-DE) with immobilized pH gradients is in turn the best approach to large-scale protein-expression screening. Spots visualization is pivotal for protein identification by mass spectrometry. Commonly used staining methods with excellent mass spectrometry compatibility are coomassie brilliant blue (CBB) or fluorescent dyes. In this study, an implementation of 'blue silver' colloidal CBB staining, characterized by high sensitivity and immediate low background, is discussed. The sensitivity of classical, colloidal and 'blue silver' CBB staining methods was compared on monodimensional and 2-DE gels. The implementation of the 'blue silver' method performs better, provided the physical state of the micelles is respected. An example of a 2-DE of human urine treated with combinatorial peptide ligand libraries demonstrates that implemented 'blue silver' can evidence the complexity of the sample.
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