Difference Gel Electrophoresis, 1st ed. 2018 Methods and Protocols Methods in Molecular Biology Series, Vol. 1664

This volume provides a comprehensive study of this key method of gel-based proteomics. Chapters in Difference Gel Electrophoresis: Methods and Protocols detail methods on principles of differential protein labeling, two-dimensional gel electrophoresis, techniques on optimized proteomic workflows, advanced mass spectrometry for protein identification, application of basic biological research, and applied biomarker discovery. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Difference Gel Electrophoresis: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Part I: Introduction

 1. Two Dimensional Gel Electrophoresis and 2D-DIGE

Paula Meleady

Part II: Difference Gel Electrophoresis

2. Comparative DIGE Proteomics

Kay Ohlendieck

3. 2D-DIGE and Fluorescence Image Analysis

4. DIGE Analysis Software and Protein Identification Approaches

Abduladim Hmmier and Paul Dowling

Part III: DIGE Applications  

5. Native DIGE: Efficient Tool to Elucidate Protein Interactomes  

Diksha Dani and Norbert Dencher

6. Comparative Two-dimensional Fluorescence Gel Electrophoresis

Doreen Ackermann and Simone König

7. DIGE-based Phosphoproteomic Analysis

Taras Stasyk and Lukas Huber

8. DIGE Saturation Labeling for Scarce Amounts of Protein from Formalin-fixed Paraffin-embedded (FFPE) Tissue

Paul Dowling

9. Comparative 3-sample DIGE Analysis of Skeletal Muscles

Kay Ohlendieck

10. DIGE Analysis of ProteoMiner™ Fractionated Serum/Plasma Samples

Sandra Murphy and Paul Dowling

Part IV: Comparative DIGE Tissue Proteomics

11. DIGE Analysis of Human Tissues

Cecilia Gelfi and Daniele Capitanio

12. DIGE Analysis of Animal Tissues

Alessio Di Luca, Ruth Hamill, Anne Maria Mullen, and Giuliano Elia

13. Rapid 2D DIGE Proteomic Analysis of Mouse Liver

Shotaro Kamata and Isao Ishii

14. Proteomic Analysis of Lung Tissue by DIGE

Jarlath Nally and Simone Schuller

15. Comparative Testis Tissue Proteomics using 2-dye versus 3-dye DIGE Analysis

Ashling Holland

16. DIGE Analysis of Fish Tissues

Joanna Nynca, Mariola Dietrich and Andrzej Ciereszko

Part V: Auxillary DIGE Methods

Sandra Murphy and Kay Ohlendieck

18. Subcellular Fractionation for DIGE-based Proteomics

19. DIGE Analysis of Immunodepleted Plasma

Paul Dowling and Kay Ohlendieck

Part VI: DIGE Verification Analysis

 20. Elucidating Cellular Metabolism and Protein Difference Data from DIGE Proteomics Experiments using Enzyme Assays

Andrew Dowd

21. Enzyme Assay Methods to Validate DIGE Proteomics Data

Andrew Dowd

22. Immunoblot Analysis of DIGE-based Proteomics

Martin Landsberger and Heinrich Brinkmeier

23. Immunofluorescence Microscopy for DIGE-based Proteomics

Rustam Mundegar, Margit Zweyer, and Dieter Swandulla

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts