2D DIGE Proteomics Service
2D gel electrophoresis involves isoelectric focusing (IEF) as 1st dimension and SDS-PAGE as 2nd dimension, separating proteins by both pI (in IEF) and molecular weight (in SDS-PAGE). 2D-DIGE proteomics analysis is an improved version of standard 2D gel electrophoresis. In addition to all features of 2D gel, 2D DIGE labels different samples with up to 3 fluorescent tags, offering significant advantages that standard 2D gel electrophoresis & other proteomics techniques cannot achieve. Please view here for comparisons of experimental procedures between 2D-DIGE and 2D gel electrophoresis.
Furthermore, our 2D DIGE proteomics services employ large format 2D gel electrophoresis, customized sample preparations and versatile study designs, offering a powerful platform for the following applications:
- 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
Advantages of 2D DIGE over 2D Gel Electrophoresis:
- Higher sensitivity: 2D DIGE uses Fluorescent labeling with a sensitivity of 0.2 ng/spot, versus the sensitivity of standard 2D gel electrophoresis with Coomassie blue staining 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: Our large 2D DIGE gel format 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 2D DIGE gel can accommodate up to 3 different samples, versus 1 sample on each 2D Gel. For the same number of samples, only 1/3 number of 2D DIGE gels is needed.
- Lower cost: Our price is based on each gel instead of each sample. Lower number of 2D DIGE gels and no need of technical replicates translates into higher quality data at a lower cost than standard 2D Gel service. Price covers from sample preparation to publication-ready data.
- Fast turn-around: Results In 5-7 Days.
2D DIGE experimental procedure:
- Sample preparation: Proteins are extracted following customized sample preparation protocols. Protein concentrations are determined by protein assay and adjusted to the desired concentration.
- Sample labeling with fluorescent dyes: Equal amount of protein extract from each sample is labeled by fluorescent dyes (size and charge matched). 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-PAGE in the 2nd dimension.
- Image acquisition: After 2D gel 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 data including the image of individual sample and overlay image of 2 or 3 samples.
- 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 are 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: Proteins are analyzed in clusters according to their functions. This is 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 for various Sample Types
2D image of jurkat cell proteome
Human Jurkat cells (pH 3-10 NL)
2D image of human PBMC proteome
Human PBMC (pH 3-10)
2D image of HEK293 cell proteome
HEK293 cell (pH 3-10 L)
Human blood T cells
2D image of human blood T cell proteome
Human blood T cell (pH 3-10)
Human blood monocytes
2D image of human blood monocyte proteome
Human blood monocyte (pH 3-10)
Human dendritic cells
2D image of human dendritic cell proteome
Human dendritic cell (pH 3-10)
Human breast cancer cell line
2D image of human breast cancer cell line MCF7
Human breast cancer cell line (pH 3-10)
Human beta-islet cells
2D image of human beta-islet cell proteome
Human beta-islet cell (pH 3-10)
2D image of human hepatocyte proteome
Human hepatocyte (pH 3-10)
2D image of human keratinocyte proteome
Human keratinocyte (pH 3-10)
2D image of human platelet proteome
Human platelet (pH 3-10 L)
Human blood cell exosome
2D image of human blood cell exosome proteome
Human blood cell exosome (pH 3-10)
2D image of human saliva proteome
Saliva (pH 4-7)
Saliva (pH 3-10)
2D image of human adult urine proteome
Urine (pH 4-7)
Urine (pH 3-10)
Human amniotic fluid
2D image of human amniotic fluid proteome
Human amniotic fluid (pH 3-10)
2D image of human adipose proteome
Human adipose proteome (pH 3-10)
Human liver tissue
2D image of human liver tissue proteome
Human liver tissue (pH 3-10)
2D image of human serum proteome. See our serum proteomics page for more info.
Serum (pH 4-7)
Serum (pH 3-10)
Human cerebrospinal fluid
2D image of human cerebrospinal fluid proteome
Human cerebrospinal fluid (pH 3-10)
Mouse & Rat Brain
2D images of mouse & rat brain proteome
Mouse brain (pH 3-10)
Rat brain (pH 3-10)
2D image of mouse oocyte proteome
Mouse oocyte (pH 3-10)
Mouse retina cells
2D image of mouse retina cell proteome
Mouse retina cell (pH 3-10)
2D image of rat tear proteome
Rat tear (pH 3-10)
2D image of drosophila proteome
Drosophila (pH 3-10 NL)
2D images of C. elegans proteome
C. elegans (pH 4-7)
C. elegans (pH 3-10)
2D image of zebrafish proteome
Zebrafish (pH 3-10 L)
2D image of E. coli proteome
E. coli (pH 3-10 NL)
E. coli (pH 3-10 NL)
2D image of salmonella proteome
Salmonella (pH 3-10 L)
2D image of bacteria-secreted proteins proteome
Bacteria-secreted proteins (pH 3-10)
2D image of yeast cell proteome
Yeast (pH 3-10 NL)
Yeast (pH 3-10 NL)
2D image of plant (N. benthamiana) proteome
N. benthamiana (pH 3-10)
2D image of rice proteome
Rice (pH 3-10)
2D image of flu vaccines from different manufacture lots
Manufacturer lot 1
Manufacturer lot 2
M1 / M2