2D DIGE Proteomics Service

2D-DIGE proteomics analysis, which labels different protein samples with up to three fluorescent tags, combined with versatile study designs, offers significant advantages over standard 2D gel 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: 3 Samples in 1 Assay. Price covers from sample preparation to publication-ready data, and complimentary consultation with your project leader
  • Fast turn-around: Results In 5-7 Days.

Main applications:

2D DIGE experimental procedure

2D DIGE & mass spectrometry full procedure
  1. Sample preparation: Proteins are extracted from cells or tissues of interest. Protein concentrations are determined by protein assay and adjusted to the desired concentration.
  2. Sample labeling with fluorescent dyes: Equal amounts of protein extract are 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.
  3. 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.
  4. 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).
  5. Image analysis: ImageQuant software is used to generate the image presentation data including the single and overlay images.
  6. 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.
  7. 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.
  8. Cluster Analysis: Proteins are analyzed in clusters according to their functions. This is a complimentary service for projects with 50 or more identified proteins.
  9. Validation: We offer 2D Western Blot services to validate the 2D DIGE data.

2D DIGE Images

Human CellsHuman Fluid/TissueLab AnimalsBacteria & Others
Jurkat cellsBreast cancer cell lineSalivaMouse & Rat brainE. coli
PBMCsBeta-isletUrineMouse oocyteSalmonella
HEK293HepatocytesAmniotic fluidMouse retina cellsBacteria-secreted
Blood T cellsKeratinocytesAdiposeRat tearYeast
Blood monocytePlateletsLiver tissueDrosophilaPlant
Dendritic cellsBlood cell exosomeSerum / PlasmaC. elegansRice
Cerebrospinal fluidZebrafishFlu vaccines

Jurkat cells

2D image of jurkat cell proteome

Human Jurkat cells (pH 3-10 NL)

2D image of jurkat cell proteome

Human PBMCs

2D image of human PBMC proteome

Human PBMC (pH 3-10)

2D image of human PBMC proteome

HEK293 cells

2D image of HEK293 cell proteome

HEK293 cell (pH 3-10 L)

2D image of HEK293 cell proteome

Human blood T cells

2D image of human blood T cell proteome

Human blood T cell (pH 3-10)

2D image of human blood T cell proteome

Human blood monocytes

2D image of human blood monocyte proteome

Human blood monocyte (pH 3-10)

2D image of human blood monocyte proteome

Human dendritic cells

2D image of human dendritic cell proteome

Human dendritic cell (pH 3-10)

2D image of human dendritic cell proteome

Human breast cancer cell line

2D image of human breast cancer cell line MCF7

Human breast cancer cell line (pH 3-10)

2D image of human breast cancer cell line MCF7

Human beta-islet cells

2D image of human beta-islet cell proteome

Human beta-islet cell (pH 3-10)

2D image of human beta-islet cell proteome

Human hepatocytes

2D image of human hepatocyte proteome

Human hepatocyte (pH 3-10)

2D image of human hepatocyte proteome

Human keratinocytes

2D image of human keratinocyte proteome

Human keratinocyte (pH 3-10)

2D image of human keratinocyte proteome

Human platelets

2D image of human platelet proteome

Human platelet (pH 3-10 L)

2D image of human platelet proteome

Human blood cell exosome

2D image of human blood cell exosome proteome

Human blood cell exosome (pH 3-10)

2D image of human blood cell exosome proteome

Human saliva

2D image of human saliva proteome

Saliva (pH 4-7)

2D image of human saliva proteome pH 4-7

Saliva (pH 3-10)

2D image of human saliva proteome pH 3-10

Human urine

2D image of human adult urine proteome

Urine (pH 4-7)

2D image of human adult urine proteome pH 4-7

Urine (pH 3-10)

2D image of human adult urine proteome pH 3-10

Human amniotic fluid

2D image of human amniotic fluid proteome

Human amniotic fluid (pH 3-10)

2D image of human amniotic fluid proteome

Human adipose

2D image of human adipose proteome

Human adipose proteome (pH 3-10)

2D image of human adipose proteome

Human liver tissue

2D image of human liver tissue proteome

Human liver tissue (pH 3-10)

2D image of human liver tissue proteome

Human serum

2D image of human serum proteome. See our serum proteomics page for more info.

Serum (pH 4-7)

2D image of human serum proteome pH 4-7

Serum (pH 3-10)

2D image of human serum proteome pH 3-10

Human cerebrospinal fluid

2D image of human cerebrospinal fluid proteome

Human cerebrospinal fluid (pH 3-10)

2D image of human cerebrospinal fluid proteome

Mouse & Rat Brain

2D images of mouse & rat brain proteome

Mouse brain (pH 3-10)

2D image of mouse brain proteome

Rat brain (pH 3-10)

2D image of rat brain proteome

Mouse oocyte

2D image of mouse oocyte proteome

Mouse oocyte (pH 3-10)

2D image of mouse oocyte proteome

Mouse retina cells

2D image of mouse retina cell proteome

Mouse retina cell (pH 3-10)

2D image of mouse retina cell proteome

Rat tear

2D image of rat tear proteome

Rat tear (pH 3-10)

2D image of rat tear proteome

Drosophila

2D image of drosophila proteome

Drosophila (pH 3-10 NL)

2D image of drosophila proteome

C. elegans

2D images of C. elegans proteome

C. elegans (pH 4-7)

2D image of C. elegans proteome pH 4-7

C. elegans (pH 3-10)

2D image of C. elegans proteome pH 3-10

Zebrafish

2D image of zebrafish proteome

Zebrafish (pH 3-10 L)

2D image of zebrafish proteome

E. coli

2D image of E. coli proteome

E. coli (pH 3-10 NL)

2D color image of E. coli proteome

E. coli (pH 3-10 NL)

2D black-white image of E. coli proteome

Salmonella

2D image of salmonella proteome

Salmonella (pH 3-10 L)

2D image of salmonella proteome

Bacteria-secreted proteins

2D image of bacteria-secreted proteins proteome

Bacteria-secreted proteins (pH 3-10)

2D image of bacteria-secreted proteins proteome

Yeast

2D image of yeast cell proteome

Yeast (pH 3-10 NL)

2D color image of yeast cell proteome

Yeast (pH 3-10 NL)

2D black-white image of yeast cell proteome

Plant

2D image of plant (N. benthamiana) proteome

N. benthamiana (pH 3-10)

2D image of N. benthamiana proteome

Rice

2D image of rice proteome

Rice (pH 3-10)

2D image of rice proteome

Flu Vaccines

2D image of flu vaccines from different manufacture lots

Manufacturer lot 1

2D image of flu vaccine from manufacturer lot 1

Manufacturer lot 2

2D image of flu vaccine from manufacturer lot 2

M1 / M2

2D overlay image of flu vaccines

2D DIGE FAQs

Frequently asked questions about 2D-DIGE advantages, process, time & cost:

Compared to 2D-gel, 2D-DIGE offers higher sensitivity, higher accuracy, fewer number of gels, faster turn around, and a lower price.

Learn the difference

The sensitivity of 2D DIGE is about 0.2 ng/spot.

2D DIGE is very accurate due to the extremely high spot resolution. Differences in protein expression as small as 10% can be detected.

Highly reproducible: identical data can be obtained on the same sample labeled with different CyDye on the same gel or cross different gels (see an example).

No

2000-5000 protein spots using pH3-10 IPG strip. If using additional IPG strips such as pH 4-7, pH 6-9 and pH 7-11, over 5000 spots can be detected.

Most customers request to start the experiment with an Analytical Gel for the following reasons:

  1. Analytical gels use about 25 µg of each sample while prep-gels use about 250 µg of each sample. The total protein load for analytical gel is about 75 µg, while total protein load for prep-gels is about 750 µg. Since Analytical Gel contains lower sample amount resulting in less ion and salt in IEF, it will provide more accurate quantitation of protein changes) and much higher image quality for publication/presentation purposes.
  2. After running the analytical gel, the in-gel data analysis will be done. Based on the analytical gel results, you can put more control or treatment samples if your desired spots are from control (usually down-regulation), or from treatment (usually up-regulation or newly induced proteins). In contrast, Prep-gel is much less likely to provide such information due to the excess amount of protein.
  3. From the analytical gel, you will know if you need to focus on a specific region (i.e. with most changes) in the next 2-D experiment by changing the pH range for IEF or the percentage of the SDS-gel (for resolving lower or higher MW protein spots). In contrast, Prep-gel is much less likely to provide such information due to the excess amount of protein.
  4. The main purpose of running Prep-gel is to obtain enough protein amount for the protein ID by Mass Spectrometry.

5-7 days for Analytical gel.

7-9 days for Prep-gel, spot picking and protein identification by mass spectrometry.

The data report contains two main parts:

  • ImageQuant analysis: gel images of individual samples and overlay of two sample images
  • DeCyder analysis: quantitative analysis and comparison of all spots between different samples.

Yes, we can send you the raw data generated by instrument software free of charge within 1 year of the completion of project. If you request raw data after 1 year, it may incur a charge based on the actual time to retrieve them from achieve.

Let’s assume a new customer from academics who is interested in finding the differential protein expression among 2-3 samples, followed by obtaining Protein ID for the top 10 spots. Please note that the prices are per gels, not per sample. Each gel can load up to 3 samples. Following would be the estimated cost:

Service Description2 samples3 samples
2D DIGE Analytical Gel$1050*$1195*
2D DIGE Preparative Gel$1200$1295
Spot Picking (up to 96 spots per gel)$250$250
10 Protein ID by Mass Spectrometry$1300$1300
Total cost$3800$4040

* Only applies for new customers who submit standard protein samples with protein concentration >5mg/ml. The discount is subject to approval from Finance. The discounted price applies for the first 2D DIGE Analytical gel. The discounted price does NOT apply for Preparative gels.