Fluorescent dual colour 2D-protein gel electrophoresis for rapid detection of differences in protein pattern with standard image analysis software

F, Von Eggeling; Gawriljuk, A; Fiedler, Wolfgang; Ernst, Günther; Claussen, U; Klose, Joachim; Römer, Irmgard

doi:10.3892/ijmm.8.4.373

Cited by 27 publications

(20 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2-D DIGE has already been employed for examining the protein profiles of various tissues and cell types including those from bacteria, 4,12,30 mouse liver, 8,31 metaplasia/cancer cells 9,10,32,33 and adult cat/kitten brain, 34 but few studies 4,12,30,33 have thus far utilised the recently introduced DeCyder BVA for quantification and analysis of protein expression. Here, we have described how 2-D DIGE may be successfully applied to human postmortem brain investigations, and how, in conjunction with BP and BVA, protein expression may be accurately quantified across individual biological samples.…”

Section: Discussionmentioning

confidence: 99%

“…2-D DIGE coupled with DeCyder DIA or the equivalent has generally been used for quantification of protein expression. [8][9][10][11] However, there are a number of disadvantages of using DIA with a study requiring analysis of individual biological samples. The primary limitation of DIA is that the software only allows for single pair-wise comparisons.…”

Section: Image Analysismentioning

confidence: 99%

See 1 more Smart Citation

Protein profiling of human postmortem brain using 2-dimensional fluorescence difference gel electrophoresis (2-D DIGE)

et al. 2004

View full text Add to dashboard Cite

Two-dimensional gel electrophoresis (2-D GE) is a key tool for comparative proteomics research. With its ability to separate complex protein mixtures with high resolution, 2-D GE is a technique commonly employed for protein profiling studies. Significant improvements have been made in 2-D GE technology with the development of two-dimensional fluorescence difference gel electrophoresis (2-D DIGE), where proteins are first labelled with one of three spectrally resolvable fluorescent cyanine dyes before being separated over first and second dimensions according to their charge and size, respectively. When used in conjunction with automated analysis packages, this multiplexing approach can accurately and reproducibly quantify protein expression for control and experimental groups. Differentially expressed proteins can be subsequently identified by mass spectrometric methods. Here, we describe the successful application and optimisation of 2-D DIGE technology for human postmortem brain studies. This technology, especially when coupled with other functional genomics approaches, such as transcriptomics and metabolomics studies, will enhance our current understanding of human disease and lead to new therapeutic and diagnostic possibilities.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Image Analysismentioning

confidence: 99%

Protein profiling of human postmortem brain using 2-dimensional fluorescence difference gel electrophoresis (2-D DIGE)

et al. 2004

View full text Add to dashboard Cite

show abstract

“…2-D DIGE has been reported to improve the speed, reproducibility, and sensitivity of 2-D PAGE-based proteomic analysis [13,[23][24][25], but not the overall drudgery associated with the technology and inability to effectively analyze high-molecular weight proteins. The 2-D DIGE concept is based on the covalent labeling of protein extracts with different fluorescent dyes, e.g., cyanine (Cy2, Cy3, or Cy5) dyes or Alexa dyes [13,25,26]. Two main protocols for 2-D DIGE are the minimal and saturated labeling and these have been described by Somiari et al [27].…”

Section: -D Difference Gel Electrophoresis (Dige)mentioning

confidence: 99%

Proteomics in human cancer research

Pastwa

Somiari

Czyż

et al. 2007

Proteomics Clinical Apps

View full text Add to dashboard Cite

Proteomics is now widely employed in the study of cancer. Many laboratories are applying the rapidly emerging technologies to elucidate the underlying mechanisms associated with cancer development, progression, and severity in addition to developing drugs and identifying patients who will benefit most from molecular targeted compounds. Various proteomic approaches are now available for protein separation and identification, and for characterization of the function and structure of candidate proteins. In spite of significant challenges that still exist, proteomics has rapidly expanded to include the discovery of novel biomarkers for early detection, diagnosis and prognostication (clinical application), and for the identification of novel drug targets (pharmaceutical application). To achieve these goals, several innovative technologies including 2-D-difference gel electrophoresis, SELDI, multidimensional protein identification technology, isotope-coded affinity tag, solid-state and suspension protein array technologies, X-ray crystallography, NMR spectroscopy, and computational methods such as comparative and de novo structure prediction and molecular dynamics simulation have evolved, and are being used in different combinations. This review provides an overview of the field of proteomics and discusses the key proteomic technologies available to researchers. It also describes some of the important challenges and highlights the current pharmaceutical and clinical applications of proteomics in human cancer research.

show abstract

“…Difference Gel Electrophoresis (DIGE) technology adds an essential quantitative component to two-dimensional gel-based proteomics to a level whereby even subtle changes in protein abundance and charge-altering post-translation modifications (such as acetylation and phosphorylation, among others) can be monitored from multiple experimental conditions with statistical confidence (49)(50)(51)(52)(53). DIGE overcomes many of the limitations commonly associated with two-dimensional gels, such as analytical (gel-to-gel) variation and limited dynamic range that can severely hamper a quantitative differential-display study.…”

Section: Quantitative Proteomicsmentioning

confidence: 99%

Mass Spectrometry-based Proteomic Profiling of Lung Cancer

Ocak¹,

Chaurand²,

Massion³

2009

Proceedings of the American Thoracic Society

View full text Add to dashboard Cite

In an effort to further our understanding of lung cancer biology and to identify new candidate biomarkers to be used in the management of lung cancer, we need to probe these tissues and biological fluids with tools that address the biology of lung cancer directly at the protein level. Proteins are responsible of the function and phenotype of cells. Cancer cells express proteins that distinguish them from normal cells. Proteomics is defined as the study of the proteome, the complete set of proteins produced by a species, using the technologies of large-scale protein separation and identification. As a result, new technologies are being developed to allow the rapid and systematic analysis of thousands of proteins. The analytical advantages of mass spectrometry (MS), including sensitivity and highthroughput, promise to make it a mainstay of novel biomarker discovery to differentiate cancer from normal cells and to predict individuals likely to develop or recur with lung cancer. In this review, we summarize the progress made in clinical proteomics as it applies to the management of lung cancer. We will focus our discussion on how MS approaches may advance the areas of early detection, response to therapy, and prognostic evaluation.Keywords: proteome; translational research; biomarkers Lung cancer is the leading cause of cancer-related death worldwide among both males and females, with more than 1 million deaths annually (1). Non-small cell lung cancer (NSCLC) accounts for about 80% of all lung cancers. Although advances have been made in diagnosis and treatment strategies in the last decade, the prognosis of NSCLC patients is poor, with a 5-year overall survival of 15 to 20% (2). This is mainly due to a lack of early diagnosis tools, with more than 60% of the patients diagnosed with advanced or metastatic disease (3) and therefore not eligible for a curative surgical resection. Lung cancer is often suspected on the basis of abnormal chest imaging and/or nonspecific symptoms. Bronchoscopy, with cytopathologic examination of bronchoalveolar lavage, endobronchial brushings and biopsies obtained from the suspect area, is in general used as an initial diagnostic tool. However, while this procedure is 100% specific for lung cancer, the sensitivity is low and ranges from 30% for small peripheral lesions to 80% for central endobronchial tumors (4). More invasive and expensive diagnostic tests are often required, delaying the diagnosis and the subsequent treatment initiation. Surgical resection offers the best chance for cure. For patients undergoing surgery, the longterm prognosis remains disappointing, with a 5-year overall survival of 50% only (5). Recent studies showed that survival of surgically resected patients with NSCLC might be improved by systemic platinum-based adjuvant chemotherapy (6, 7), but which patients might benefit from this treatment cannot be determined accurately.To improve lung cancer management and survival, there is a great need to develop screening and early diagnosis strategies that are sensitive,...

show abstract

Fluorescent dual colour 2D-protein gel electrophoresis for rapid detection of differences in protein pattern with standard image analysis software

Cited by 27 publications

References 0 publications

Protein profiling of human postmortem brain using 2-dimensional fluorescence difference gel electrophoresis (2-D DIGE)

Protein profiling of human postmortem brain using 2-dimensional fluorescence difference gel electrophoresis (2-D DIGE)

Proteomics in human cancer research

Mass Spectrometry-based Proteomic Profiling of Lung Cancer

Contact Info

Product

Resources

About