Difference gel electrophoresis (DIGE) : methods and protocols / edited by Rainer Cramer, Reiner Westermeier.

Contributor(s): Cramer, Rainer | Westermeier, Reiner
Material type: TextTextSeries: Methods in molecular biology (Clifton, N.J.): 854. 1064-3745.Publisher: New York : Humana Press, ©2012Description: 1 online resource (xiii, 401 pages)Content type: text Media type: computer Carrier type: online resourceISBN: 9781617795732; 1617795739; 1617795720; 9781617795725Subject(s): Gel electrophoresis -- Laboratory manuals | Proteomics -- Laboratory manuals | Electrophoresis, Gel, Two-Dimensional | Publication Formats | Biochemistry | Genomics | Investigative Techniques | Electrophoresis, Polyacrylamide Gel | Genetics | Analytical, Diagnostic and Therapeutic Techniques and Equipment | Biological Science Disciplines | Publication Characteristics | Computational Biology | Electrophoresis | Chemistry | Chemistry Techniques, Analytical | Natural Science Disciplines | Biology | Electrochemical Techniques | Disciplines and Occupations | Two-Dimensional Difference Gel Electrophoresis | Proteomics | Laboratory Manuals | Methods | Gel electrophoresis | Proteomics | Life sciences | Proteomics | Biochemistry | Protein Science | eiwitten | proteins | eiwitexpressieanalyse | biologische technieken | biological techniques | Biological Techniques | Biologische techniekenGenre/Form: Electronic books. | Laboratory manuals. Additional physical formats: Printed edition:: No titleDDC classification: 543/.4 LOC classification: QP519.9.G42 | D54 2012Online resources: Click here to access online
Contents:
Fundamentals -- DIGE : past and future / Jonathan S. Minden -- Basics of 2D DIGE / Phil Beckett -- Multifluorescence 2D gel imaging and image analysis / Ingo Vormbrock, Sonja Hartwig, and Stefan Lehr -- Assessing signal-to-noise in quantitative proteomics : multivariate statistical analysis in DIGE experiments / David B. Friedman -- Analysis of proteins using DIGE and MALDI mass spectrometry / Witold M. Winnik [and others] -- Synthesis and validation of cyanine-based dyes for DIGE / Michael E. Jung [and others] -- Methods -- 2D DIGE saturation labeling for minute sample amounts / Georg J. Arnold and Thomas Frohlich -- Proteomic analysis of redox-dependent changes using cysteine-labeling 2D DIGE / Hong-Lin Chan, John Sinclair, and John F. Timms -- Analysis of protein posttranslational modifications using DIGE-based proteomics / Robert M. DeKroon [and others] -- Comparative analyses of protein complexes by blue native DIGE / Katrin Peters and Hans-Peter Braun -- 2D DIGE analysis of protein extracts from muscle tissue / Cecilia Gelfi and Sara De Palma -- Combination of highly efficient hexapeptide ligand library-based sample preparation with 2D DIGE for the analysis of the hidden human serum/plasma proteome / Sonja Hartwig and Stefan Lehr -- 2D DIGE analysis of serum after fractionation by proteominertm beads / Cynthia Liang, Gek San Tan, and Maxey C.M. Chung -- Study design in DIGE-based biomarker discovery / Alexandra Graf and Rudolf Oehler -- Comparative 2D DIGE analysis of the depleted serum proteome for biomarker discovery / Megan Penno, Matthias Ernst, and Peter Hoffmann -- Applications in clinical proteomics -- Differential gel-based proteomic approach for cancer biomarker discovery using human plasma / Keun Na [and others] -- 2D DIGE for the analysis of RAMOS cells subproteomes / Marisol Fernandez and Juan Pablo Albar -- Application of saturation labeling in lung cancer proteomics / Gereon Poschmann [and others] -- Proteomic profiling of the epithelial-mesenchymal transition using 2D DIGE / Rommel A. Mathias, Hong Ji, and Richard J. Simpson -- Method for protein subfractionation of cardiovascular tissues before DIGE analysis / Athanasios Didangelos, Xiaoke Yin, and Manual Mayr -- Application of DIGE and mass spectrometry in the study of type 2 diabetes mellitus mouse models / Celia Smith, Davinia Mills, and Rainer Cramer -- Evaluating the efficacy of subcellular fractionation of blast cells using live cell labeling and 2D DIGE / Yin Ying Ho [and others] -- Applications in animal, plant, and microbial proteomics -- DIGE analysis of plant tissue proteomes using a phenolic protein extraction method / Christina Rode [and others] -- Native DIGE of fluorescent plant protein complexes / Veronika Reisinger and Lutz Andreas Eichacker -- Overview of 2D DIGE analysis of marine (environmental) bacteria / Ralf Rabus -- Application of 2D DIGE in animal proteomics / Ingrid Miller.
Summary: Protein analysis is increasingly becoming a cornerstone in deciphering the molecular mechanisms of life. Proteomics, the large-scale and high-sensitivity analysis of proteins, is already pivotal to the new life sciences such as Systems Biology and Systems Medicine. Proteomics, however, relies heavily on the past and future advances of protein purification and analysis methods. DIGE, being able to quantify proteins in their intact form, is one of a few methods that can facilitate this type of analysis and still provide the protein isoforms in an MS-compatible state for further identification and characterization with high analytical sensitivity. Differential Gel Electrophoresis: Methods and Protocols introduces the concept of DIGE and its advantages in quantitative protein analysis. It provides detailed protocols and important notes on the practical aspects of DIGE with both generic and specific applications in the various areas of Quantitative Proteomics. Divided into four concise sections, this detailed volume opens with the basics of DIGE, the technique and its practical details with a focus on the planning of a DIGE experiment and its data analysis. The next section introduces various DIGE methods from those employed by scientists world-wide to more novel methods, providing a glance at what is on the horizon in the DIGE world. The volume closes with an overview of the wide range of DIGE applications from Clinical Proteomics to Animal, Plant, and Microbial Proteomics applications. Written in the highly successful Methods in Molecular Biology series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Differential Gel Electrophoresis: Methods and Protocols can be used by novices with some background in biochemistry or molecular biology as well as by experts in Proteomics who would like to deepen their understanding of DIGE and its employment in many hyphenations and application areas. With its many protocols, applications, and methodological variants, it is also a unique reference for all who seek fundamental details on the working principle of DIGE and ideas for possible future uses of DIGE in novel analytical approaches.
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Includes index.

Part I Fundamentals -- DIGE : past and future / Jonathan S. Minden -- Basics of 2D DIGE / Phil Beckett -- Multifluorescence 2D gel imaging and image analysis / Ingo Vormbrock, Sonja Hartwig, and Stefan Lehr -- Assessing signal-to-noise in quantitative proteomics : multivariate statistical analysis in DIGE experiments / David B. Friedman -- Analysis of proteins using DIGE and MALDI mass spectrometry / Witold M. Winnik [and others] -- Synthesis and validation of cyanine-based dyes for DIGE / Michael E. Jung [and others] -- Part II Methods -- 2D DIGE saturation labeling for minute sample amounts / Georg J. Arnold and Thomas Frohlich -- Proteomic analysis of redox-dependent changes using cysteine-labeling 2D DIGE / Hong-Lin Chan, John Sinclair, and John F. Timms -- Analysis of protein posttranslational modifications using DIGE-based proteomics / Robert M. DeKroon [and others] -- Comparative analyses of protein complexes by blue native DIGE / Katrin Peters and Hans-Peter Braun -- 2D DIGE analysis of protein extracts from muscle tissue / Cecilia Gelfi and Sara De Palma -- Combination of highly efficient hexapeptide ligand library-based sample preparation with 2D DIGE for the analysis of the hidden human serum/plasma proteome / Sonja Hartwig and Stefan Lehr -- 2D DIGE analysis of serum after fractionation by proteominertm beads / Cynthia Liang, Gek San Tan, and Maxey C.M. Chung -- Study design in DIGE-based biomarker discovery / Alexandra Graf and Rudolf Oehler -- Comparative 2D DIGE analysis of the depleted serum proteome for biomarker discovery / Megan Penno, Matthias Ernst, and Peter Hoffmann -- Part III Applications in clinical proteomics -- Differential gel-based proteomic approach for cancer biomarker discovery using human plasma / Keun Na [and others] -- 2D DIGE for the analysis of RAMOS cells subproteomes / Marisol Fernandez and Juan Pablo Albar -- Application of saturation labeling in lung cancer proteomics / Gereon Poschmann [and others] -- Proteomic profiling of the epithelial-mesenchymal transition using 2D DIGE / Rommel A. Mathias, Hong Ji, and Richard J. Simpson -- Method for protein subfractionation of cardiovascular tissues before DIGE analysis / Athanasios Didangelos, Xiaoke Yin, and Manual Mayr -- Application of DIGE and mass spectrometry in the study of type 2 diabetes mellitus mouse models / Celia Smith, Davinia Mills, and Rainer Cramer -- Evaluating the efficacy of subcellular fractionation of blast cells using live cell labeling and 2D DIGE / Yin Ying Ho [and others] -- Part IV Applications in animal, plant, and microbial proteomics -- DIGE analysis of plant tissue proteomes using a phenolic protein extraction method / Christina Rode [and others] -- Native DIGE of fluorescent plant protein complexes / Veronika Reisinger and Lutz Andreas Eichacker -- Overview of 2D DIGE analysis of marine (environmental) bacteria / Ralf Rabus -- Application of 2D DIGE in animal proteomics / Ingrid Miller.

Protein analysis is increasingly becoming a cornerstone in deciphering the molecular mechanisms of life. Proteomics, the large-scale and high-sensitivity analysis of proteins, is already pivotal to the new life sciences such as Systems Biology and Systems Medicine. Proteomics, however, relies heavily on the past and future advances of protein purification and analysis methods. DIGE, being able to quantify proteins in their intact form, is one of a few methods that can facilitate this type of analysis and still provide the protein isoforms in an MS-compatible state for further identification and characterization with high analytical sensitivity. Differential Gel Electrophoresis: Methods and Protocols introduces the concept of DIGE and its advantages in quantitative protein analysis. It provides detailed protocols and important notes on the practical aspects of DIGE with both generic and specific applications in the various areas of Quantitative Proteomics. Divided into four concise sections, this detailed volume opens with the basics of DIGE, the technique and its practical details with a focus on the planning of a DIGE experiment and its data analysis. The next section introduces various DIGE methods from those employed by scientists world-wide to more novel methods, providing a glance at what is on the horizon in the DIGE world. The volume closes with an overview of the wide range of DIGE applications from Clinical Proteomics to Animal, Plant, and Microbial Proteomics applications. Written in the highly successful Methods in Molecular Biology series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Differential Gel Electrophoresis: Methods and Protocols can be used by novices with some background in biochemistry or molecular biology as well as by experts in Proteomics who would like to deepen their understanding of DIGE and its employment in many hyphenations and application areas. With its many protocols, applications, and methodological variants, it is also a unique reference for all who seek fundamental details on the working principle of DIGE and ideas for possible future uses of DIGE in novel analytical approaches.

Includes bibliographical references and index.

English.

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