Journal
NATURE PROTOCOLS
Volume 1, Issue 4, Pages 1732-1742Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2006.256
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Funding
- NIDA NIH HHS [R01 DA013772-03] Funding Source: Medline
- NIMH NIH HHS [R01 MH074313, R01 MH074313-02] Funding Source: Medline
- NATIONAL INSTITUTE OF MENTAL HEALTH [R01MH074313] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE ON DRUG ABUSE [R01DA013772] Funding Source: NIH RePORTER
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Quantitative proteomics is the workhorse of the modern proteomics initiative. The gel-based and MuDPIT approaches have facilitated vital advances in the measurement of protein expression alterations in normal and disease phenotypic states. The methodological advance in two-dimensional gel electrophoresis (2DGE) has been the multiplexing fluorescent two-dimensional fluorescence difference gel electrophoresis (2D-DIGE). 2D-DIGE is based on direct labeling of lysine groups on proteins with cyanine CyDye DIGE Fluor minimal dyes before isoelectric focusing, enabling the labeling of 2-3 samples with different dyes and electrophoresis of all the samples on the same 2D gel. This capability minimizes spot pattern variability and the number of gels in an experiment while providing simple, accurate and reproducible spot matching. This protocol can be completed in 3-5 weeks depending on the sample size of the experiment and the level of expertise of the investigator.
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