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

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

Content Provider	Springer Nature Link
Author	Gauci, Victoria J. Wright, Elise P. Coorssen, Jens R.
Copyright Year	2010
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.
Starting Page	3
Ending Page	29
Page Count	27
File Format	PDF
ISSN	18646158
Journal	Journal of Chemical Biology
Volume Number	4
Issue Number	1
e-ISSN	18646166
Language	English
Publisher	Springer-Verlag
Publisher Date	2010-06-19
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	Protein staining Fluorescence Coomassie Sypro Ruby 2D gel electrophoresis Analytical proteomics Biochemistry Pharmacology/Toxicology Cell Biology Biophysics and Biological Physics Physical Chemistry
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Biophysics

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