Two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) is a quantitative proteomics technique developed from traditional two-way electrophoresis. 2D-DIGE separates mixed proteins based on the same principle as two-way electrophoresis, using the difference in isoelectric point and molecular weight of proteins to separate proteins. The fluorescent dyes used in DIGE are Cy2, Cy3, and Cy5, which can react with the lysine side chain amino groups of proteins to label the proteins, and the isoelectric point and molecular weight of the labeled proteins are not affected. After mixing the labeled proteins in equal amounts and performing bi-directional electrophoresis, the differences between gels can be eliminated by matching cy2 internal standards, and the changes in protein expression are reflected by the different fluorescence intensities of cy3 and cy5.

What We Offer

Lifeasible provides SDS-PAGE and 2D-DIGE electrophoresis services combined with an advanced mass spectrometry platform and nanoLC-MS/MS nanoliter chromatography to provide one-stop plant proteome characterization and quantification services to researchers worldwide. This service applies to a wide range of samples and there is no requirement for sample status, either liquid or solid samples. Lifeasible will provide you with a detailed technical report after the experimental results, including the experimental steps, relevant experimental parameters, gel plots, mass spectrometry images, raw data, and professional 2D-DIGE quantitative proteomics analysis results.

Service Flow

Technical Characteristics

• Low experimental consumption without the need for expensive isotope labels as internal standards.
• Less manipulation of the sample, thus bringing it closest to its original state.
• Independence from sample conditions, overcoming the drawbacks of labeled quantitative techniques in performing analysis on multiple samples.
• High requirements for stability and reproducibility of experimental operations, requiring at least three biological replicates.

Reference

1. Kathryn S. L., et al. "Methods of quantitative proteomics and their application to plant organelle characterization." Journal of Experimental Botany, 2006, 57(7): 1493-1499.

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For research or industrial use.

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