Skip to main content

Application of Proteomics in Cancer Gene Profiling: Two-Dimensional Difference in Gel Electrophoresis (2D-DIGE)

  • Protocol
  • First Online:
Cancer Gene Profiling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 576))

Summary

In the post-genomic era, proteomic strategies are at the forefront of cancer research. By studying the complement of all expressed genes, proteomics aims to provide knowledge of biomarkers indicative of the physiological state of cancer cells at a specific time, enabling screening, early diagnosis, monitoring the course of cancer development/progression, and gauging the efficacy and safety of novel therapeutic agents. Onco-proteomics thus has the ability to revolutionise oncology practice by delivering highly selective and individualised clinical care.

One of the proteomic techniques, two-dimensional (2D) difference in gel electrophoresis (DIGE) enables simultaneous examination and comparison of multiple samples using cyanine dyes to label amino acid residues that are then separated based on charge and mass. This technique reduces variability, improves reproducibility, and allows easier quantitation when compared with traditional 2D polyacrylamide gel electrophoresis (PAGE). These advantages combined with universal availability makes 2D-DIGE a first method of choice in cancer proteome analysis of diverse specimens, including tissues, cell lines, blood, and other body fluids.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Pastwa, E., Somiari, S. B., Czyz, M. and Somiari, R. I. (2007) Proteomics in human cancer research. Proteomics Clin Appl. 1(1), 4–17.

    Article  CAS  Google Scholar 

  2. O’Farrell, P. H. (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 250, 4007–4021.

    PubMed  Google Scholar 

  3. Unlu, M., Morgan, M. E. and Minden, J. S. (1997) Difference gel electrophoresis: a single gel method for detecting changes in protein extracts. Electrophoresis. 18, 2071–2077.

    Article  CAS  PubMed  Google Scholar 

  4. Marouga, R., David, S. and Hawkins, E. (2005) The development of the DIGE system: 2D fluorescence difference gel analysis technology. Anal Bioanal Chem. 382, 669–678.

    Article  CAS  PubMed  Google Scholar 

  5. Tonge, R., Shaw, J., Middleton, B., Rowlinson, R., Rayner, S., Young, J., et al. (2001) Validation and development of fluorescence two-dimensional differential gel electrophoresis proteomics technology. Proteomics. 1, 377–396.

    Article  CAS  PubMed  Google Scholar 

  6. DeSouza, L., Diehl, G., Rodrigues, M. J., Guo, J., Romaschin, A. D., Colgan, T. J., et al. (2005) Search for cancer markers from endometrial tissues using differentially labeled tags iTRAQ and cICAT with multidimensional liquid chromatography and tandem mass spectrometry. J Proteome Res. 4, 377–386.

    Article  CAS  PubMed  Google Scholar 

  7. Ryu, O. H., Atkinson, J. C., Hoehn, G. T., Illei, G. G. and Hart, T. C. (2006) Identification of parotid salivary biomarkers in Sjogren’s syndrome by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry and two-dimensional difference gel electrophoresis. Rheumatology (Oxford). 45, 1077–1086.

    Article  CAS  Google Scholar 

  8. Shaw, M. M. and Riederer, B. M. (2003) Sample preparation for two-dimensional gel electrophoresis. Proteomics. 3, 1408–1417.

    Article  CAS  PubMed  Google Scholar 

  9. Orenes-Pinero, E., Corton, M., Gonzalez-Peramato, P., Algaba, F., Casal, I., Serrano, A., et al. (2007) Searching urinary tumor markers for bladder cancer using a two-dimensional differential gel electrophoresis (2D-DIGE) approach. J Proteome Res. 6, 4440–4448.

    Article  CAS  PubMed  Google Scholar 

  10. Lee, I. N., Chen, C. H., Sheu, J. C., Lee, H. S., Huang, G. T., Yu, C. Y., et al. (2005) Identification of human hepatocellular carcinoma-related biomarkers by two-dimensional difference gel electrophoresis and mass spectrometry. J Proteome Res. 4, 2062–2069.

    Article  CAS  PubMed  Google Scholar 

  11. Katayama, M., Nakano, H., Ishiuchi, A., Wu, W., Oshima, R., Sakurai, J., et al. (2006) Protein pattern difference in the colon cancer cell lines examined by two-dimensional differential in-gel electrophoresis and mass spectrometry. Surg Today. 36, 1085–1093.

    Article  CAS  PubMed  Google Scholar 

  12. Jin, T., Hu, L. S., Chang, M., Wu, J., Winblad, B. and Zhu, J. (2007) Proteomic identification of potential protein markers in cerebrospinal fluid of GBS patients. Eur J Neurol. 14, 563–568.

    Article  CAS  PubMed  Google Scholar 

  13. Kakisaka, T., Kondo, T., Okano, T., Fujii, K., Honda, K., Endo, M., et al. (2007) Plasma proteomics of pancreatic cancer patients by multi-dimensional liquid chromatography and two-dimensional difference gel electrophoresis (2D-DIGE): up-regulation of leucine-rich alpha-2-glycoprotein in pancreatic cancer. J Chromatogr B Analyt Technol Biomed Life Sci. 852, 257–267.

    Article  CAS  PubMed  Google Scholar 

  14. Gharbi, S., Gaffney, P., Yang, A., Zvelebil, M. J., Cramer, R., Waterfield, M. D., et al. (2002) Evaluation of two-dimensional differential gel electrophoresis for proteomic expression analysis of a model breast cancer cell system. Mol Cell Proteomics. 1, 91–98.

    Article  CAS  PubMed  Google Scholar 

  15. Thiede, B., Hohenwarter, W., Krah, A., Mattow, J., Schmid, M., Schmidt, F., et al. (2005) Peptide mass fingerprinting. Methods. 35, 237–247.

    Article  CAS  PubMed  Google Scholar 

  16. Aebersold, R. and Goodlett, D. R. (2001) Mass spectrometry in proteomics. Chem Rev. 101, 269–295.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2009 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Hariharan, D., Weeks, M.E., Crnogorac-Jurcevic, T. (2009). Application of Proteomics in Cancer Gene Profiling: Two-Dimensional Difference in Gel Electrophoresis (2D-DIGE). In: Grützmann, R., Pilarsky, C. (eds) Cancer Gene Profiling. Methods in Molecular Biology, vol 576. Humana Press. https://doi.org/10.1007/978-1-59745-545-9_11

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-545-9_11

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-934115-76-3

  • Online ISBN: 978-1-59745-545-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics