Comparative and Semi-Quantitative Proteomics Using 2D Gel Electrophoresis (2D-DIGE) Technology
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ITSI-Biosciences utilizes validated sample preparation kits/protocols, robotic spot pickers/digesters, DeCyder difference-in-gel analysis software and advanced mass spectrometers to offer an integrated 2D-DIGE – MS/MS proteomics solution. Proteomic analysis usually starts from sample preparation, to analyses by mass spectrometry (LC-MS/MS) and protein identification and annotation. Data obtained after 2D-DIGE analysis may include protein identity, pI, molecular weight and post translational modifications.
ITSI-Biosciences has provided 2D-DIGE analysis and proteomics services to NIH, industry, biotechnology companies, pharmaceutical companies, Research Institutes and Universities. Our more than 15 years of experience with 2D-DIGE analysis, proteomics, mass spectrometry and protein expression profiling enables us to offer reproducible picture perfect gels at a competitive price and guaranteed faster turnaround time. We have also developed a variety of complementary 2D-DIGE kits to streamline and standardize protein isolation, protein quantitation, protein qualification and the 2D-DIGE analysis step, thereby reducing the overall time and cost of performing 2D-DIGE analysis.
2D-DIGE offers all the advantages of the classical 2D-PAGE, and most importantly overcomes the inherent disadvantage of gel-to-gel variation and reproducibility problem inherent with the traditional 2D-PAGE approach. 2D-DIGE is presently the most versatile gel-based approach for comparative and semi-quantitative proteomics. Distinct fluorescent tags e.g. Cy 3, 5 and 2 are used to label protein samples and a universal internal standard prior to 1st/2nd dimension electrophoresis. An automated software program is used to detect, quantify and annotate differentially expressed proteins. 2D-DIGE analysis can be applied to protein expression profiling and comparative proteomics by using DeCyder software to identify proteins with statistically-significant difference in expression. This provides unique opportunities for researches in fields including cancer, neuroscience, vision, cardiovascular, Alzheimer, Parkinson, etc, to find important proteins associated with the condition. 2D-DIGE – LC/MS/MS analysis can also be applied to identify post translationally modified proteins (PTM) that are important in cell signaling and protein trafficking.
| S/N | CAT NO. | NAME | DESCRIPTION | UNIT COST |
|---|---|---|---|---|
| 1 | SP1 | Sample Preparation 1 | Homogenization, protein quantitation using ToPA Bradford protein assay kit. | Per sample |
| 2 | SP2 | Sample Preparation 2 | Includes, precipitation, buffer exchange, protein quantitation, etc. and the use of ToPI-DIGE, ToPREP and ToPA kits. | Per sample |
| 3 | SP3 | Sample Preparation 3 | Depletion of abundant protein e.g. bovine or human serum albumin using ITSI ASK-Column kit. | Per sample |
| 4 | SP4 | Sample Preparation 4 | Depletion of abundant protein e.g. bovine or human serum albumin using ITSI ASK-Solvent kit. | Per sample |
| 5 | SP5 | Sample Preparation 5 | Depletion IgY Column Depletion of 14 abundant proteins from serum /plasma using IgY column. | Per sample |
| 6 | SP9 | Sample Preparation 9 | Urine protein isolation and concentration. Service includes Isolation, partial purification, and concentration of urine proteins using ITSI Urine Protein Isolation kit. | Per sample |
| 7 | S30 | 2D-DIGE | Two- Dimensional Difference in-Gel Electrophoresis (2D- DIGE). Quantitative and comparative proteomics using 2D-DIGE. Services include: • Experimental design, • Sample preparation to isolate total proteins using ToPI- DIGE Protein Isolation Kit, • Fractionation, quantitation and qualification of proteins, • 2D-DIGE analysis using Cy2, Cy3 and Cy5, scanning of gels using a DIGE- enabled digital scanner and • Biological variation analysis of gel images to identify differentially expressed proteins with statistically significant difference. | 2 – 10 Samples |
| 8 | S40P | Robotic spot picking | Selection and picking of fluorescent labels protein spots from 2D-DIGE gels using fully automated ETTAN spot picking robot. | 1-96 spots |
| 9 | S40D | Robotic spot digestion | In gel digestion of protein spots picked from 2D-DIGE gels using a fully automated ETTAN spot digestion robot. | 1-96 spots |
| 10 | S50.60 | LC60-MS/MS | Peptide sequencing and protein identification by liquid chromatography – tandem mass spectrometry of pure protein samples. Services include: • Tandem mass spectrometry analysis using nano-LC/MS/MS, • Database search and • Identified protein annotation – 60 min run. | Per sample |
