2D DIGE Proteomics Service
2D-DIGE proteomics analysis, which labels different protein samples with up to three fluorescent tags, offers significant advantages over conventional 2D-PAGE and other proteomics platforms:
- Higher sensitivity: Fluorescent dyes have a sensitivity of 0.2 ng/spot, versus the sensitivity of coomassie at 100 ng/spot or silver staining at 1 ng/spot
- Higher accuracy: The extremely high spot resolution enables accurate spot quantitation. Differences in protein expression as small as 10% can be detected
- Higher reproducibility: Nearly identical data was obtained on the same sample labeled with different CyDye on the same gel or cross different gels, thus eliminating the need of running technical replicates
- Broader spectrum: Each gel can resolve ~5000 protein spots, allowing a wide dynamic range detection/quantitation of low abundant proteins, large proteins and small peptides
- Fewer number of gels: Each gel can accommodate up to 3 different samples
- Lower cost: The price is based on the number of gels, not the number of samples. Price covers from sample preparation to publication-ready data, and complimentary consultation with your project leader
- Fast turn-around: 1 week
- Identifying therapeutic or diagnostic markers in different diseases
- Identifying key regulators in major biological pathways
- Studying post-translational modifications such as phosphorylation, glycosylation, etc.
- Determining HCP antibody coverage and detecting HCP contaminants
2D DIGE experimental procedure
- Sample preparation: Proteins are extracted from cells or tissues of interest. Protein concentrations are determined by protein assay and adjusted to the desired concentration.
- Sample labeling with fluorescent dyes: Equal amount of protein extract was labeled by fluorescent dyes (size and charge matched), and the spectrally resolvable dyes enable simultaneous co-separation and analysis of up to 3 samples on a single multiplexed gel.
- 2D gel electrophoresis: Up to 3 samples can be simultaneously separated on a single 2D gel, with isoelectric focusing (IEF) in the 1st dimension and SDS polyacrylamide gel electrophoresis (SDS-PAGE) in the 2nd dimension.
- Image acquisition: After electrophoresis, the gel is scanned using a Typhoon image scanner. Each scan reveals one of the CyDye signals (Cy2, Cy3 and Cy5).
- Image analysis: ImageQuant software is used to generate the image presentation data including the single and overlay images.
- Quantitative analysis: Comparative analysis of all spots using DeCyder “in-gel” or “cross-gel” analysis software. The protein expression ratios between different samples or different groups of samples will be generated.
- Automated spot picking: Following the spot picking design using DeCyder software, protein spots of interest can be automatically picked from the 2D gel with the Ettan Spot Picker, followed by Protein Identification by Mass Spectrometry.
- Cluster Analysis: A complimentary service for projects with 50 or more identified proteins.
- Validation: We offer 2D Western Blot services to validate the 2D DIGE data.
2D DIGE Images
|Table of Contents|
|Yeast||Mouse & Rat Brain|
2D images of human saliva proteome
Saliva pH 4-7
Saliva pH 3-10
2D images of human adult urine proteome
2D images of E. coli Proteome (pH 3-10 NL)
2D image of yeast (pH 3-10 NL)
2D image of drosophila proteome (pH 3-10 NL)
2D image of plant (N. benthamiana) proteome (pH 4-7)
2D images of C. elegans proteome
C. elegans: pH 4-7
C. elegans: pH 3-10
2D image of zebrafish proteome (pH 3-10L)
Mouse & Rat Brain
2D images of mouse & rat brain proteome
Mouse Brain: pH 3-10
Rat Brain: pH 3-10
2D image of rat tear proteome