Kurien / Scofield Electrophoretic Separation of Proteins

Methods and Protocols
Erscheinungsjahr 2018
ISBN: 978-1-4939-8793-1
Verlag: Springer US
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 1855, 523 Seiten, eBook

Reihe: Methods in Molecular Biology

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.

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Kurien, Biji T.

Scofield, R. Hal

Inhaltsverzeichnis

How It All Began: A Personal History of Gel-Electrophoresis.- Introduction to Protein Electrophoresis.- Concentrating Proteins by Salt, Polyethylene Glycol, Solvent, SDS Precipitation, Three Phase Partitioning, Dialysis, Centrifugation, Ultrafiltration, Lyophilisation, Affinity Chromatography, Immunoprecipitation or Increased Temperature for Protein Isolation, Drug Interaction, and Proteomic and Peptidomic Evaluation.- Lysis Buffer Choices are Key Considerations to Ensure Effective Sample Solubilization for Protein Electrophoresis.- The Cydex Blue Assay: A One Step Protein Assay for Samples Prior to SDS Electrophoresis.- Cellulose-Acetate Electrophoresis of Hemoglobin.- Native Polyacrylamide Gels.- Isoelectric Focusing on Non-Denaturing Gels.- Determination of protein Molecular weights on SDS-PAGE.- Two-Dimensional Gel Electrophoresis by Glass Tube-Based IEF and SDS-PAGE.- Cationic Electrophoresis.- Two-Dimensional Gel Electrophoresis with Immobilized pH Gradients.- SARCOSYL-PAGE: Optimized Protocols for the Separation and Immunological Detection of PEGylated Proteins.- Tricine-SDS-PAGE.- Analysis of Protein Glycation using Phenylboronate Acrylamide Gel Electrophoresis.- Immunofixation Electrophoresis for Identification of Proteins and Specific Antibodies.- Electrophoretic Separation of Very Large Molecular Weight Proteins in SDS-Agarose.- Increase in Local Protein Concentration by Field-Inversion Gel Electrophoresis.- Two-Dimensional Difference Gel Electrophoresis.- Immunoelectrophoresis: A Method with Many Faces.- Tris-Acetate Polyacrylamide Gradient Gels for the Simultaneous Electrophoretic Analysis of Proteins if Very High and Low Molecular Mass.- Diagnoal Electrophoresis for Detection of Proteins Involved in Disulfide Bonds.- Identification of Proteins on Archived 2D Gels.- Two-Dimensional Gel Electrophoresis: Vertical Isoelectric Focusing.- A Foodomics Approach: CE-MS for Comparative Metabolomics of Colon Cancer Cells Treated with Dietary Polyphenols.- Characterization of New Cyclic D, L – a – Alternate Amino Acid Peptides by Capillary Electrophoresis Coupled to Electrospray Ionization Mass Spectrometry.- “Microchip Electrophoresis”, with Respect to “Profiling of Aß Peptides in the Cerebrospinal Fluid of Patients with Alzheier’s Disease”.- Measuring Low-Picomolar Apparent Binding Affinities by Minigel Electrophoretic Mobility Shift.- Identification of Proteins Interacting with Single Nucleotide Polymorphisms (SNPs) by DNA Pull-Down Assay.- Horizontal Agarose Gel Mobility Shift Assay for Protein-RNA Complexes.- Applications of Immobilized Metal affinity Electrophoresis.- Isoelectric Focusing in Agarose Gel for Detection of Oligoclonal Bands in Cerebrospinal and Other Biological Fluids.- A Combined Free Flow Electrophoresis and DIGE Approach to Compare Proteins in Complex Biological Samples.- SDS PAGE for 35S Immunoprecipitation and Immunoprecipitation Western Blotting.- A Multichannel Gel Electrophoresis and Continuous Fraction Collection Apparatus for High-Throughput Protein Separation and Characterization.- Cell Surface Protein Biotinylation for SDS-PAGE Analysis.- Isolation of Proteins from Polyacrylamide Gels.- Continuous Elution SDS-PAGE with a Modified Standard Gel Apparatus to Separate and Isolate an Array of Proteins from Complex Mixtures.- Protein Extraction from Gels: A Brief Review.- Gel Absorption-Based Sample Preparation Method for Shotgun Analysis of Membrane Proteome.- Ultra-Rapid Sodium Dodecyl Sulfate Polyacrylamide Mini-Gel Electrophoresis.- A Brief Review of Other Notable Electrophoretic Methods.- Single-Cell High Resolution Detection and Quantification of Protein Isoforms Differing by a Single Charge Unit.- Artifacts and Common Errors in Protein Gel Electrophoresis.