Protein Electrophoresis

Each chapter in this Methods in Molecular Biology(TM) book details a specific electrophoretic variant, so that scientists can perform new techniques without difficulty. Includes step-by-step protocols, troubleshooting advice and tips on avoiding pitfalls.

Proteins are the functional units of the cellular machinery and they provide significant information regarding the molecular basis of health and disease. Therefore, techniques to separate and isolate the various proteins are critical to studying and understanding their functional characteristics. One of the widely used techniques for this purpose is electrophoresis. In Protein Electrophoresis: Methods and Protocols, contributions from experts in the field have been collected in order to provide practical guidelines to this complex study. Each chapter outlines a specific electrophoretic variant in detail so that laboratory scientists may perform a technique new to their lab without difficulty. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and accessible, Protein Electrophoresis: Methods and Protocols seeks to serve laboratory scientists with well-honed, detailed methodologies in an effort to further our knowledge of this essential field.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Contenu

How It All Began -- A Personal History of Gel-Electrophoresis.- Introduction to Protein Electrophoresis.- Protein Quantification Methods to Determine Protein Concentration Prior to Electrophoresis.- Protein Solubilization: Attend to the Choice of Lysis Buffer.- Native Polyacrylamide Gels.- Cationic Electrophoresis.- SARCOSYL-PAGE: A New Electrophoretic Method for the Separation and Immunological Detection of PEGylated Proteins.- Tricine-SDS-PAGE.- Analysis of Protein Glycation using Phenylboronate Acrylamide Gel Electrophoresis.- Protein Electrophoresis in Agarose Gels for Separating High Molecular Weight Proteins.- Increase in Local Protein Concentration by Field-Inversion Gel Electrophoresis.- A Combined Free Flow Electrophoresis and DIGE Approach to Compare Proteins in Complex Biological Samples.- Immunofixation Electrophoresis for Identification of Proteins and Specific Antibodies.- Microchip Electrophoresis, with respect to Profiling of A Peptides in the Cerebrospinal Fluid of Patients with Alzheimer's Disease.- A Foodomics Approach: CE-MS for Comparative Metabolomics of Colon Cancer Cells Treated with Dietary Polyphenols.- Peptide Fractionation by SDS-Free Polyacrylamide Gel Electrophoresis for Proteomic Analysis via DF-PAGE.- Tris-Acetate Polyacrylamide Gradient Gels for the Simultaneous Electrophoretic Analysis of Proteins of Very High and Low Molecular Mass.- SDS PAGE for 35S Immunoprecipitation and Immunoprecipitation WesternBlotting.- Two Dimensional Gel Electrophoresis: Vertical Isoelectric Focusing.- Isoelectric Focusing in Agarose Gel for Detection of Oligoclonal Bands in Cerebrospinal and Other Biological Fluids.- Native Flat-Bed Isoelectric Focusing for Determining Antibody Clonotype Distribution.- Two-Dimensional Gel Electrophoresis: Glass Tube-Based IEF Followed by SDS-PAGE.- One-Dimensional and Two-Dimensional Immobilized Metal Affinity Electrophoresis.- Two-Dimensional Difference Gel Electrophoresis.- Protein Identification on Archived 2-D Gels.- Diagonal Electrophoresis for the Detection of Protein Disulfides.- Two-Dimensional Blue Native/SDS Gel Electrophoresis of Multiprotein Complexes.- Immunoelectrophoresis: A Method with Many Faces.- Cell Surface Protein Biotinylation for SDS PAGE Analysis.- A Multichannel Gel Electrophoresis and Continuous Fraction Collection Apparatus for High-Throughput Protein Separation and Characterization.- Gel Absorption-Based Sample Preparation Method for Shotgun Analysis of Membrane Proteome.- Interactions of Hemoglobin in Live Red Blood Cells Measured by the Electrophoresis Release Test.- Extraction of Proteins from Gels - A Brief Review.- A New Algorithm for the Simulation of SDS 2D-PAGE Datasets.- Isolation of Proteins from Polyacrylamide Gels.- Gel Drying Methods.- Long-Term, Buffer-Less Wet Gel Storage in Non-Sealed Polyethylene Bags.- A Brief Review of Other NotableElectrophoretic Methods.- Protein Stains and Applications.- Coomassie-Brilliant Blue Staining of Polyacrylamide Gels.- Accelerated Coomassie Blue Protein Staining and Destaining with Application of Heat.- Silver Staining Techniques of Polyacrylamide Gels.- Revisit of Imidazole-Zinc Reverse Stain for Protein Polyacrylamide Gel Electrophoresis.- Counterion Dye Staining of Proteins in One- and Two-Dimensional Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis and Tryptic Gel Digestion of Stained Protein for Mass Spectrometry.- TEMED Enhanced Photoluminescent Imaging of Human Serum Proteins by Quantum Dots after PAGE.- Microwave Assisted Protein Staining, De-Staining and In-Gel / In-Solution Digestion of Proteins.- Electrophoretic Separation and Detection of Metalloproteins by X-ray Fluorescence Mapping.- Fluorescent Staining of Gels.- A Single-Step Simultaneous Protein Staining Procedure for Polyacrylamide Gels and Nitrocellulose Membranes by Alta During Western Blot Analysis.- Detection of Glycoproteins in Polyacrylamide Gels Using Pro-Q Emerald 300 Dye, a Fluorescent Periodate Schiff-Base Stain.- Spicy SDS-PAGE Gels: Curcumin/Turmeric as an Environment-Friendly Protein Stain.- In-Gel Staining of Proteins in Native Poly Acryl Amide Gel Electrophoresis Using meso-Tetrakis(4-sulfonato phenyl)porphyrin.- Rapid Detection of Proteins in Polyacrylamide Electrophoresis Gels with Direct Red 81 and Amido Black.- Zymographic Assay of Oxidases Using Peroxidase or HeminEntrapped in Polyacrylamide Gels.- Coomassie Brilliant Blue Removal/Disposal from Gel Destain and Used Gel Stain in an Environment-Friendly Manner.- A Brief Review of Other Notable Protein Detection Methods on Acrylamide Gels.- Ultra-Sensitive Protein Detection and Imaging: Comparison of Lumitein™, ProteoSilver™, SYPRO® Ruby, and Coomassie® Brilliant Blue Gel Stains.- Common Artifacts and Mistakes Made in Electrophoresis.