Electrophoretic Separation of Proteins: Methods and Protocols

Description

This volume expands upon Protein Electrophoresis (2012) and provides readers with easy-to-follow and reproducible methods to study electrophoresis. The chapters in this book cover topics such as the Cydex Blue assay; cellulose-acetate electrophoresis of hemoglobin; cationic electrophoresis; tricine-SDS-Page; identification of proteins on archived 2-D gels; cell surface protein biotinylation of SDS-PAGE analysis; and artifacts and common errors in protein gel electrophoresis. 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. Practical and thorough, Electrophoretic Separation of Proteins: Methods and Protocols is a valuable resource for researchers who are interested in learning and experimenting with this field. Front Matter ....Pages i-xv How It All Began: A Personal History of Gel Electrophoresis (Oliver Smithies)....Pages 1-21 Introduction to Protein Electrophoresis (Pothur R. Srinivas)....Pages 23-29 Measuring Protein Concentration with Absorbance, Lowry, Bradford Coomassie Blue, or the Smith Bicinchoninic Acid Assay Before Electrophoresis (J. P. Dean Goldring)....Pages 31-39 Concentrating Proteins by Salt, Polyethylene Glycol, Solvent, SDS Precipitation, Three-Phase Partitioning, Dialysis, Centrifugation, Ultrafiltration, Lyophilization, Affinity Chromatography, Immunoprecipitation or Increased Temperature for Protein Isolation, Drug Interaction, and Proteomic and Peptidomic Evaluation (J. P. Dean Goldring)....Pages 41-59 Lysis Buffer Choices Are Key Considerations to Ensure Effective Sample Solubilization for Protein Electrophoresis (Ewa I. Miskiewicz, Daniel J. MacPhee)....Pages 61-72 The Cydex Blue Assay: A One-Step Protein Assay for Samples Prior to SDS Electrophoresis (Thierry Rabilloud)....Pages 73-80 Cellulose Acetate Electrophoresis of Hemoglobin (Ramesh Kumar, Wilbert A. Derbigny)....Pages 81-85 Native Polyacrylamide Gels (Claudia Arndt, Stefanie Koristka, Anja Feldmann, Michael Bachmann)....Pages 87-91 Isoelectric Focusing on Non-Denaturing Flatbed Gels (Biji T. Kurien, R. Hal Scofield)....Pages 93-100 Determination of Protein Molecular Weights on SDS-PAGE (Hiroyuki Matsumoto, Hisao Haniu, Naoka Komori)....Pages 101-105 Two-Dimensional Gel Electrophoresis by Glass Tube-Based IEF and SDS-PAGE (Hiroyuki Matsumoto, Hisao Haniu, Biji T. Kurien, Naoka Komori)....Pages 107-113 Cationic Electrophoresis (Engelbert Buxbaum)....Pages 115-124 Two-Dimensional Gel Electrophoresis with Immobilized pH Gradients (Bre’Ana Byrd, Huyen Tran)....Pages 125-129 SARCOSYL—PAGE: Optimized Protocols for the Separation and Immunological Detection of PEGylated Proteins (Christian Reichel, Günter Gmeiner, Philipp Reihlen, Mario Thevis, Wilhelm Schänzer)....Pages 131-149 Tricine-SDS-PAGE (Syed R. Haider, Helen J. Reid, Barry L. Sharp)....Pages 151-160 Analysis of Protein Glycation Using Phenylboronate Acrylamide Gel Electrophoresis (Marta P. Pereira Morais, Omar Kassaar, Stephen E. Flower, Robert J. Williams, Tony D. James, Jean M. H. van den Elsen)....Pages 161-175 Immunofixation Electrophoresis for Identification of Proteins and Specific Antibodies (Gyorgy Csako)....Pages 177-201 Electrophoretic Separation of Very Large Molecular Weight Proteins in SDS Agarose (Marion L. Greaser, Chad M. Warren)....Pages 203-210 Increase in Local Protein Concentration by Field-Inversion Gel Electrophoresis (Henghang Tsai, Hon-Chiu Eastwood Leung)....Pages 211-227 Two-Dimensional Difference Gel Electrophoresis (Malachi Blundon, Vinitha Ganesan, Brendan Redler, Phu T. Van, Jonathan S. Minden)....Pages 229-247 Immunoelectrophoresis: A Method with Many Faces (Gyorgy Csako)....Pages 249-268 Tris-Acetate Polyacrylamide Gradient Gels for the Simultaneous Electrophoretic Analysis of Proteins of Very High and Low Molecular Mass (Monica Cubillos-Rojas, Fabiola Amair-Pinedo, Irantzu Tato, Ramon Bartrons, Francesc Ventura, Jose Luis Rosa)....Pages 269-277 Diagonal Electrophoresis for the Detection of Proteins Involved in Disulfide Bonds (Ronald Saraswat, Brian McDonagh)....Pages 279-286 Identification of Proteins on Archived 2D Gels (Hiroyuki Matsumoto, Nobuaki Takemori, Naoka Komori)....Pages 287-289 Two-Dimensional Gel Electrophoresis: Vertical Isoelectric Focusing (Yaser Dorri)....Pages 291-302 A Foodomics Approach: CE-MS for Comparative Metabolomics of Colon Cancer Cells Treated with Dietary Polyphenols (Mustafa Çelebier, Clara Ibáñez, Carolina Simó, Alejandro Cifuentes)....Pages 303-313 Characterization of New Cyclic d,l-α-Alternate Amino Acid Peptides by Capillary Electrophoresis Coupled to Electrospray Ionization Mass Spectrometry (María Dámaris Cortez-Díaz, Fanny d’Orlyé, Anne Varenne)....Pages 315-326 “Microchip Electrophoresis,” with Respect to “Profiling of Aβ Peptides in the Cerebrospinal Fluid of Patients with Alzheimer’s Disease” (Mohamad Reza Mohamadi, Romain Verpillot, Myriam Taverna, Markus Otto, Jean-Louis Viovy)....Pages 327-340 Measuring Low-Picomolar Apparent Binding Affinities by Minigel Electrophoretic Mobility Shift (Karen A. Lewis, Sarah E. Altschuler, Deborah S. Wuttke)....Pages 341-354 Identification of Proteins Interacting with Single Nucleotide Polymorphisms (SNPs) by DNA Pull-Down Assay (Bhupinder Singh, Swapan K. Nath)....Pages 355-362 Horizontal Agarose Gel Mobility Shift Assay for Protein-RNA Complexes (Jennifer A. Ream, L. Kevin Lewis, Karen A. Lewis)....Pages 363-370 Applications of Immobilized Metal Affinity Electrophoresis (Bao-Shiang Lee, Lasanthi P. Jayathilaka, Jin-Sheng Huang, Shalini Gupta)....Pages 371-385 Isoelectric Focusing in Agarose Gel for Detection of Oligoclonal Bands in Cerebrospinal and Other Biological Fluids (Gyorgy Csako)....Pages 387-401 A Combined Free-Flow Electrophoresis and DIGE Approach to Compare Proteins in Complex Biological Samples (Kim Y. C. Fung, Chris Cursaro, Tanya Lewanowitsch, Leah Cosgrove, Peter Hoffmann)....Pages 403-415 SDS-PAGE for 35S Immunoprecipitation and Immunoprecipitation Western Blotting (Edward P. Trieu, Ira N. Targoff)....Pages 417-436 A Multichannel Gel Electrophoresis and Continuous Fraction Collection Apparatus for High-Throughput Protein Separation and Characterization (Ming Dong, Megan Choi, Mark D. Biggin, Jian Jin)....Pages 437-448 Cell Surface Protein Biotinylation for SDS-PAGE Analysis (Giuliano Elia)....Pages 449-459 Isolation of Proteins from Polyacrylamide Gels (Stefanie Koristka, Claudia Arndt, Ralf Bergmann, Michael Bachmann)....Pages 461-465 Continuous Elution SDS-PAGE with a Modified Standard Gel Apparatus to Separate and Isolate an Array of Proteins from Complex Mixtures (Robert G. E. Krause, J. P. Dean Goldring)....Pages 467-478 Protein Extraction from Gels: A Brief Review (Biji T. Kurien, Rachna Aggarwal, R. Hal Scofield)....Pages 479-482 Gel Absorption-Based Sample Preparation Method for Shotgun Analysis of Membrane Proteome (Xianchun Wang, Songping Liang)....Pages 483-490 Ultrarapid Sodium Dodecyl Sulfate Polyacrylamide Mini-Gel Electrophoresis (Rohit Thomas, Biji T. Kurien)....Pages 491-494 A Brief Review of Other Notable Electrophoretic Methods (James D. Fesmire)....Pages 495-499 Single-Cell High-Resolution Detection and Quantification of Protein Isoforms Differing by a Single Charge Unit (Kristi A. Koelsch)....Pages 501-509 Artifacts and Common Errors in Protein Gel Electrophoresis (Biji T. Kurien, R. Hal Scofield)....Pages 511-518 Back Matter ....Pages 519-523