Two-dimensional gel electrophoresis, abbreviated as 2-DE, is a powerful and widely used tool that uses gel electrophoresis to analyze mixtures of proteins. While both isoelectric focusing and SDS-PAGE are powerful techniques, 2D electrophoresis is a clever combination of the two methods. Proteins are first separated by their isoelectric point (pl) in isoelectric focusing, followed by a second separation through SDS-PAGE that resolves proteins based on molecular weight. Every protein spot on a 2D electrophoresis can be eluted and identified via high-throughput mass spectrometry. Therefore, 2D electrophoresis is particularly useful to compare protein profiles between different tissues, conditions, or between control and treated samples.

Overview of 2D Electrophoresis Service

At Creative Proteomics, we can provide reliable 2D electrophoresis service according to standard operating procedures (SOPs). With this service, complex mixtures consisting of thousands of different proteins can be separated and their relative amount can then be determined.

Figure 1. The workflow of 2D electrophoresis.

• Sample preparation. Samples need to be converted to a physicochemical state, and the procedures of sample preparation are dependent on the sample type.
• First separation by isoelectric focusing. A protein mixture is loaded at the basic end of the pH gradient gel. After applying an electric field, proteins with a positive net charge will migrate toward the cathode and those with a negative net charge will migrate toward the anode.
• Second dimension separation by SDS-PAGE. This method generally involves three steps, including gel preparation, the equilibrium of the immobilized pH gradient stripes in SDS buffer, and electrophoresis.
• Visualization of results. We use multiple methods (such as blue staining and silver staining) for visualization of proteins.
• Further analysis. Protein can be separated and information such as molecular weight, pl, and the amount can be determined.

Sample Requirement

We accept diverse samples, including but not limited to

• Purified proteins
• Frozen cell pellets or tissues
• Cell (such as blood cells) or tissue
• Serum / plasma / urine
• Cell cultures
• Fermentation cultures
• Microorganisms
• Plants

Advantages of 2D Electrophoresis

• High-throughput. 2D electrophoresis can accurately analyze thousands of proteins in a single run.
• High resolution. This technology resolves proteins according to both pI and molecular mass, and enables the characterization of proteins with posttranslational modifications that affect their charge state.
• Various computer-based tools are available. We have tools such as SameSpots, Delta2D, ImageMaster, which can be used for detection and quantification of protein spots.
• Cost-efficient and affordable. While mass spectrometers represent a significant investment and require experience staff, 2D electrophoresis is relatively inexpensive.
• Flexibility. We work closely with you to design the optimal experimental scheme to meet your research objectives on budget.

Applications of 2D Electrophoresis

• Protein cataloguing
• Protein species and isoform distribution analysis
• Protein quantification
• Posttranslational modification studies
• Quality control of proteins and antibody
• Host cell protein analysis
• Downstream analysis such as western blotting
• Developmental genetics, disease etiology, biomarker and target discovery

Based on professional knowledge and experienced staff, Creative Proteomics provides the high-resolution and high-throughput 2-DE service for protein analysis. This service can provide direct visualization of changes in protein profiles and post-translational modifications. It also helps the discovery and detection of biomarkers, as well as drug discovery. As every project has different requirements, please contact our specialists to discuss your specific needs.