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.
Our 2D-DIGE based comparative proteomics services includes, experimental design, Sample preparation to isolate total proteins using ToPI-DIGE protein isolation Kit, pre-fractionation, quantitation and qualification of proteins, 2D-DIGE analysis, scanning of gels using a Typhoon DIGE-enabled digital scanner, Biological Variation Analysis (BVA) of spots detected to identify differentially expressed proteins with statistically significant difference, robotic spot picking of candidate proteins, robotic in-gel digestion of candidate proteins with trypsin or another enzyme. The enzyme digested proteins are identified by Tandem Mass Spectrometry and annotated.
We offer the following services:
|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:||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:||Per sample|