Establishment of “one-piece” large-gel 2-DE for high-resolution analysis of small amounts of sample using difference gel electrophoresis saturation labelling

Sitek, B.; Sipos, B.; Pfeiffer, K.; Grzendowski, M.; Poschmann, G.; Hawranke, E.; Köper, K.; Klöppel, G.; Meyer, H. E.; Stühler, K.
May 2008
Analytical & Bioanalytical Chemistry;May2008, Vol. 391 Issue 1, p361
Academic Journal
Large-gel two-dimensional gel electrophoresis (2-DE) is the method of choice for high-resolution proteome analysis of complex protein mixtures. Until now, however, the advantages of large 2-DE in combination with multiplexed fluorescence dye protein labelling has been complicated by the separate handling and analysis of the second-dimension gels. Therefore, we adapted the large 2-DE procedure allowing us to run “one-piece” large 2-DE gels (40 cm × 30 cm) in the second dimension for high resolution proteome analysis. Here, we show that in combination with fluorescence dye protein saturation labelling “one-piece” large 2-DE enables analysis of small amounts of sample (3 µg protein) for high-resolution proteome analysis.


Related Articles

  • Current Progress in Sample Preparation for Two-Dimensional Electrophoresis in Proteomics. Zhen Cai; Jen-Fu Chiu; Qing-Yu He // Current Proteomics;Dec2005, Vol. 2 Issue 4, p259 

    Two dimensional polyacrylamide gel electrophoresis (2DE) has been a core technology of current proteomics for its high resolution and ability to detect proteins with post-translational modifications. This technology has been widely applied in numerous biomedical research projects including...

  • Cardiovascular Proteomics. Vivanco, Fernando; Dard�, Ver�nica M.; De la Cuesta, Fernando; Barderas, Mar�a G. // Current Proteomics;Oct2006, Vol. 3 Issue 3, p147 

    Cardiovascular diseases are among the leading cause of morbidity and mortality in Western societies and developing countries. The ability to investigate the complete proteome provides a critical tool toward elucidating the complex and multifactorial basis of cardiovascular biology, especially...

  • Biomarker Discovery in Clinical Proteomics: Strategies for Exposing Low Abundant Proteins. Wang, Yu; Seneviratne, Jayampath // Current Proteomics;Jul2008, Vol. 5 Issue 2, p104 

    Disease specific proteins are highly valuable as clinical biomarkers, which can be used in early diagnosis, monitoring disease progressions and evaluating therapies. The identification and characterization of protein biomarkers in different physiological and pathological sources of interest...

  • Proteomics Mapping of Cord Blood Identifies Haptoglobin "Switch-On" Pattern as Biomarker of Early-Onset Neonatal Sepsis in Preterm Newborns. Buhimschi, Catalin S.; Bhandari, Vineet; Dulay, Antonette T.; Nayeri, Unzila A.; Razeq, Sonya S. Abdel-; Pettker, Christian M.; Thung, Stephen; Guomao Zhao; Han, Yiping W.; Bizzarro, Matthew; Buhimschi, Irina A. // PLoS ONE;2011, Vol. 6 Issue 10, p1 

    Background: Intra-amniotic infection and/or inflammation (IAI) are important causes of preterm birth and early-onset neonatal sepsis (EONS). A prompt and accurate diagnosis of EONS is critical for improved neonatal outcomes. We sought to explore the cord blood proteome and identify biomarkers...

  • Out with the old? Schaffer, Catherine // Drug Discovery & Development;Dec2007, Vol. 10 Issue 12, p44 

    The article discusses ways to improve the two-dimensional polyacrylamide gel electrophoresis (2D PAGE) technology. The tool is considered as a reliable technique with proven track record for identification and qualitative analysis of proteins. It covers a complex protein sample resolution and...

  • Web-Based Tools for the Interpretation of Chain-Like Protein Spot Patterns on Two-Dimensional Gels. H. Maurer, Martin // Current Proteomics;Apr2012, Vol. 9 Issue 1, p18 

    Two-dimensional gel electrophoresis is a standard tool of proteomic analysis, consisting of first dimensional protein separation according to the respective protein's isoelectric point, and the second dimensional separation according to the protein's molecular weight. In these gels, 'chain-like'...

  • The untiring search for the most complete proteome representation: reviewing the methods. De Souza, Daniel Martins; Oliveira, Bruno M.; Castro-Dias, Erich; Winck, Flavia V.; Horiuchi, Ricardo S. O.; Baldasso, Paulo A.; Caetano, Hugo T.; Dayane Pires, Natália Katauchi; Marangoni, Sergio; Novello, José C. // Briefings in Functional Genomics & Proteomics;Jul2008, Vol. 7 Issue 4, p312 

    Proteomic research has proved valuable for understanding the molecular mechanisms of biological processes, as well as in the search for biomarkers for a variety of diseases which lack a molecular diagnostic. While several new approaches are being developed, two-dimensional (2-DE) gel...

  • Assessment of the 2-D Gel-Based Proteomics Application of Clinically Archived Formalin-Fixed Paraffin Embedded Tissues. Davalieva, Katarina; Kiprijanovska, Sanja; Polenakovic, Momir // Protein Journal;Apr2014, Vol. 33 Issue 2, p135 

    Hospital tissue repositories possess a vast and valuable supply of disease samples with matched retrospective clinical information. Detection and characterization of disease biomarkers in formalin-fixed paraffin-embedded (FFPE) tissues will greatly aid the understanding of the diseases...

  • Differential plasma proteome profiles of mild versus severe β-thalassemia/Hb E. Hatairaktham, Suneerat; Srisawat, Chatchawan; Siritanaratkul, Noppadol; Chiangjong, Wararat; Fucharoen, Suthat; Thongboonkerd, Visith; Kalpravidh, Ruchaneekorn // Annals of Hematology;Mar2013, Vol. 92 Issue 3, p365 

    The severity of thalassemia is currently classified based on clinical manifestations and multiple tests. In the present study, we performed a plasma proteome analysis to identify differentially expressed proteins compared between normal subjects and patients with mild and severe forms of...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics