Comparative and Semi-Quantitative Proteomics Using 2D Gel Electrophoresis (2D-DIGE) Technology
ITSIBIO »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 |
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Citations
Browse through a list of scientific publications and research papers involving our 2D-DIGE Services.
42. Perturbation of Iron Metabolism by Cisplatin through Inhibition of Iron Regulatory Protein 2.
Miyazawa, Masaki 2, Alexander R. Bogdan, Yoshiaki Tsuji 3. Cell Chemical Biology, November 15, 2018, Volume 26, Issue 1, 85 – 97.e4. doi: 10.1016/j.chembiol.2018.10.009
41. Identification of Protein Markers in Patients Infected with Plasmodium knowlesi, Plasmodium falciparum and Plasmodium vivax.
Alan Kang-Wai Mu, Int J Mol Sci. 2014 Nov 3;15(11):19952-61. doi: 10.3390/ijms151119952.
40. AMP-activated protein kinase mediates insulin-like and lipo-mobilising effects of β-glucan-rich polysaccharides isolated from Pleurotus sajor-caju (Fr.), Singer mushroom, in 3T3-L1 cells.
39. A colorimetric method for monitoring tryptic digestion prior to shotgun proteomics.
Somiari RI, Renganathan K, Russell S, Wolfe S, Mayko F, Somiari SB.Int J Proteomics. 2014;2014:125482. doi: 10.1155/2014/125482.
38. Wortmannin potentiates the combined effect of etoposide and cisplatin in human glioma cells.
Pastwa E, Poplawski T, Lewandowska U, Somiari SB, Blasiak J, Somiari RI. Int J Biochem Cell Biol. 2014 Aug;53:423-31.
37. DNA aptamers against exon v10 of CD44 inhibit breast cancer cell migration.
Iida J, Clancy R, Dorchak J, Somiari RI, Somiari S, Cutler ML, Mural RJ, Shriver CD. PLoS One. 2014 Feb 19;9(2):e88712.
36. A Novel Type of Hemocytes Localizing Melanization with High-Spreading Behavior in Mythimna Separata.
Yoshiaki Kato, Tatsuhiro Yoshida, Ken Miura, Toshiharu Tanaka, Yutaka Nakamatsu, Masanori Ochiai, Archives of Insect Biochemistry and Physiology 2014 Jul DOI: 10.1002/arch.21173
35. Identification of STAT2 serine 287 as a novel regulatory phosphorylation site in type I interferon-induced cellular responses.
H. C. Steen, S. Nogusa, R. J. Thapa, S.H. Basagoudanavar, A. Gill, S. Merali, C. A. Barrero, S. Balachandran, A. M. Gamero, The Journal of Biological Chemistry
34. COPD is associated with production of autoantibodies to a broad spectrum of self-antigens, correlative with disease phenotype.
T. A. Packard, Q.Z. Li, G.P. Cosgrove, R.P. Bowler, J.C. Cambier Immunol Res (2013) 55:48–57, September 2012. Springer Science+Business Media
33. Acute mitochondrial dysfunction after blast exposure: potential role of mitochondrial glutamate oxaloacetate transaminase.
P. Arun, R. Abu-Taleb, S. Oguntayo, Y. Wang, M. Valiyaveettil, J. Long, M.P. Nambiar Journal of Neurotrauma
32. Modulation of hearing related proteins in the brain and inner ear following repeated blast exposures.
P. Arun, M. Valiyaveettil1, L. Biggemann, Y. Alamneh, Y. Wei, S. Oguntayo, Y. Wang, J.B. Long, M.P. Nambiar1 … Interventional Medicine & Applied Science vol.4 2012 October 1
31. Exploration of the binding proteins of perfluorooctane sulfonate by a T7 phage display screen.
Y Miyano, S Tsukuda, I Sakimoto, R Takeuchi, Satomi Shimuraa, N Takahashia,T Kusayanagia,Y Takakusagia … Journal of Proteomics-Elsevier Inc. 2012 May 15
30. Beta-Catenin Phosphorylated at Threonine 120 Antagonizes Generation of Active Beta-Catenin by Spatial Localization in trans-Golgi Network.
Cheng Du1, Chuanyou Zhang1, Zhuo Li1, Md. Helal Uddin Biswas1, K. C. Balaji2.PLos ONE. 2012 April 12; Volume 7, Issue 4
29. How the Surface Nanostructure of Polyethylene Affects Protein Assembly and Orientation.
James D. Bortner, Arunangshu Das, Todd M. Umstead, Williard M. Freeman, Richard Somiari, Cesar Aliaga, David S. Phelps, . ACS Nano. 03/2011; 5(4).
28. Method for releasing reducing Glycan by Ammonium Salt.
Nishimura, Shin-ichiro, Miura, Yoshiaki. Free Patents online. 2011 November 10, patent:20110275108
27. Senescence Marker Protein 30 (SMP30) Expression in Eukaryotic Cells: Existence of Multiple Species and Membrane Localization.
Peethambaran Arun, Vineela Aleti, Kalpana Parikh, Veeraswamy Manne, Nageswararao Chilukuri. PLos ONE. 2011 February; Volume 6, Issue 2
26. The effect of selenium enrichment on baker’s yeast proteome.
K. El-Bayoumy, A. Dasa, S. Russell, S. Wolfe, R. Jordan, K. Renganathan, T.P. Loughranb, R. Somiari. Journal of Proteomics-Elsevier Inc. 2011 October 21;.10.1016
25. Repeated Blast Exposures Lead to Changes in Hearing Related Gene Expression in Mice Brain.
P. Arun, Y. Wang, Y. Wei, S. Oguntayo, L. Biggemann, M. P. Nambiar. RTA.nato.Int. 2011.
24. Effect of Immunohistochemistry on Molecular Analysis of Tissue Samples.
M. A. Tangrea, S. Mukherjee, B. Gao, S P. Markey, Q. Du, M. Armani M. S. Kreitman, A.M. Rosenberg, B.S. Wallis, F. C. Eberle, F.C. Duncan, J.C. Hanson, R.F. Chuaqui, J.Rodriguez-Canales, M.R. Emmert-Buck Journal of Histochemistry & Cytochemistry. 2011 February 24.
23. Differential profile analysis of urinary cytokines in patients with overactive bladder.
G.Ghoniem., N. Faruqui., M. Elmissiry. Int. Urogynecol J Springer. 2011 April13; 22:953-961
22. Polyphenols Activate Nrf2 in Astrocytes Via H2O2, Semi-Quinones and Quinones.
H. Erlanka, A. Elmanna, R. Kohenb, J. Kanner Free Radical Biology and Medicine-Elsevier Inc. 2011 September 28;10.1016
21. Sample Preparation for 2D Electrophoresis and CE-Based Proteomics .
J. M. Nagy, A. Lipka, F. Pereira, N. Marlin, S. Hassard Springer Science+Buisness Media (google book) 2011.
20. STR Analysis of Human DNA Samples After Dry-State Ambient Temperature Storage in Geneplates.
Richard I. Somiari, E. Adebiyi, L. Ukachukwu, I. Mba, F. A. Anthony, A. I. Onuaha, M.Brainard, S.Lubert, C. Larson, S. Russell, N. Bharathan, S.B. Somiari. The Open Forensic Science Journal, 2011, 4, 30-35, November 23, 2011.
19. Selenium-Responsive Proteins in the Sera of Selenium-Enriched Yeast-Supplemented Healthy American and Caucasian Men.
R. Sinha., I. Sinha., N. Facompreruqui., M. Elmissiry. Journal of Biomedical Science. 2009 September;35(3):559-67.
18. Down-regulation of 14-3-3 isoforms and annexin A5 proteins in lung adenocarcinoma induced by the tobacco-specific nitrosamine NNK in the A/J mouse revealed by proteomic analysis.
Bortner JD Jr, Das A, Umstead TM, Freeman WM, Somiari R, Aliaga C, Phelps DS, El-Bayoumy K. J Proteome Res. 2009 Aug;8(8):4050-61. doi: 10.1021/pr900406g
17. Characterization of membranous and cytoplasmic EGFR expression in human normal renal cortex and renal cell carcinoma.
16. Proteomics of rat prostate lobes treated with 2-N-Hydroxylamino-1-methyl-6-phenylimidazo [4,5-b] pyridine, 5alpha-dihydrotestosterone, individually and in combination.
Boyiri, T., Somiari, RI., Russell, S., EL-Bayoumy, K. Int. J Oncol. 2009 Sep;35(3):559-67
15. Circulating MMP2 and MMP9 in breast cancer – potential role in classification of patients into low risk, benign disease and breast cancer categories.
Somiari, S., Somiari, RI., Heckman, C., Olsen, C., Jordan R., Russell, S., Shriver, C., Cancer. 2006 Sep15;119(6):1403-11.
14. Plasma activity of matrix metalloproteinase 2 and 9 in breast disease.
Somiari, S., Craig, S., Heckman, C., Jordan R., Arciero, C., Hooke, J., Olsen, C., Garguilo, G., Somiari, RI. Cancer Lett. 2006 Feb20;223(1):98-107
11. Laser assisted microdissection in proteomic analyses.
Ellsworth DL, Russell S, Deyarmin B, Sullivan A, Brzeski H, Somiari RI, Shriver CD . In Walker J. (Editor) The Proteomics Protocols Handbook. Humana Press Inc., NJ, USA. 2005;pp. 59-66.
10. Large-format two dimensional polyacrylamide gel electrophoresis.
Brzeski, H., Sullivan, A., Somiari, R., & Shriver, C. In Walker J. (Editor) The Proteomics Protocols Handbook. Humana Press Inc., NJ, USA. 2005;pp. 119-132.
9. Serum or Plasma Sample Preparation for Two-Dimensional Gel Electrophoresis.
Sullivan, A., Brzeski, H., Somiari, R., & Shriver, C. In Walker J. (Editor) The Proteomics Protocols Handbook. Humana Press Inc., NJ, USA. 2005; pp. 49-54.
8. Anti-Sm and anti-RNP antibodies.
P. MIGLIORINI, C. BALDINI, V. ROCCHI, & S. BOMBARDIERI Autoimmunity, February 2005; 38(1): 47–54
5. High-throughput proteomic analysis of human infiltrating ductal carcinoma of the breast.
Somiari RI, Sullivan A, Russell S, Somiari S, Hu H, Jordan R, George A, Katenhusen R, Buchowiecka A, Arciero C, Brzeski H, Hooke J, Shriver C. Proteomics. 2003 Oct;3(10):1863-73.
Laser capture microdissection of paraffin-embedded tissues.
Ellsworth DL, Shriver CD, Ellsworth RE, Deyarmin B, Somiari RI. Biotechniques. 2003 Jan;34(1):42-4, 46
7. Global search for chromosomal abnormalities in infiltrating ductal carcinoma of the breast using array-comparative genomic hybridization.
Somiari SB, Shriver CD, He J, Parikh K, Jordan R, Hooke J, Hu H, Deyarmin B, Lubert S, Malicki L, Heckman C, Somiari RI. Cancer Genet Cytogenet. 2004 Dec;155(2):108-18.
High-throughput loss of heterozygosity mapping in 26 commonly deleted regions in breast cancer.
Ellsworth RE, Ellsworth DL, Lubert SM, Hooke J, Somiari RI, Shriver CD. Cancer Epidemiol Biomarkers Prev. 2003 Sep;12(9):915-9
6. Albumin depletion method for improved plasma glycoprotein analysis by two-dimensional difference gel electrophoresis.
Brzeski H, Katenhusen R, Sullivan A, Russell S, George A, Somiari RI, Shriver C. Biotechniques 2003 Dec;35(6):1128-32.
Difference In-Gel Electrophoresis in a High-Throughput Environment.
Somiari RI. Russell S, Somiari SB, Sullivan AG, Ellsworth, DL, Brzeski, H, Shriver, CD In Walker J. (Editor) The Proteomics Protocols Handbook. Humana Press Inc., NJ, USA. 2005;pp. 223 – 237.
Proteomics of breast carcinoma.
Somiari RI, Somiari S, Russell S, Shriver CD. J Chromatogr B Analyt Technol Biomed Life Sci. 2005 Feb 5;815(1-2):215-25.
