Quantitative proteomics : assessing the spectrum of in-gel protein detection methods

Victoria J. Gauci, Elise P. Wright, Jens R. Coorssen

    Research output: Contribution to journalArticlepeer-review

    77 Citations (Scopus)

    Abstract

    Proteomics research relies heavily on visualization methods for detection of proteins separated by polyacrylamide gel electrophoresis. Commonly used staining approaches involve colorimetric dyes such as Coomassie Brilliant Blue, fluorescent dyes including Sypro Ruby, newly developed reactive fluorophores, as well as a plethora of others. The most desired characteristic in selecting one stain over another is sensitivity, but this is far from the only important parameter. This review evaluates protein detection methods in terms of their quantitative attributes, including limit of detection (i.e., sensitivity), linear dynamic range, inter-protein variability, capacity for spot detection after 2D gel electrophoresis, and compatibility with subsequent mass spectrometric analyses. Unfortunately, many of these quantitative criteria are not routinely or consistently addressed by most of the studies published to date. We would urge more rigorous routine characterization of stains and detection methodologies as a critical approach to systematically improving these critically important tools for quantitative proteomics. In addition, substantial improvements in detection technology, particularly over the last decade or so, emphasize the need to consider renewed characterization of existing stains; the quantitative stains we need, or at least the chemistries required for their future development, may well already exist.
    Original languageEnglish
    Pages (from-to)3-29
    Number of pages27
    JournalJournal of Chemical Biology
    Volume4
    Issue number1
    DOIs
    Publication statusPublished - 2011

    Fingerprint

    Dive into the research topics of 'Quantitative proteomics : assessing the spectrum of in-gel protein detection methods'. Together they form a unique fingerprint.

    Cite this