Western Blotting

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts

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

Autorentext
Biji Kurien, PhD Associate Professor of Research Oklahoma Medical Research Foundation (OMRF) Arthritis & Clinical Immunology Research Program 825 NE 13th St. MS24 Oklahoma City, OK 73104 Tel: (405) 271-8000 ext 35204 Email: biji-kurien@ouhsc.edu Robert H. Scofield, MD Arthritis & Clinical Immunology Research Program, Oklahoma Medical Research Foundation 825 N.E. 13th Street. MS 38 Oklahoma City, OK 73104 Phone: (405) 271-7061 Fax: (405) 271-7063 E-mail: Hal-Scofield@omrf.org

Inhalt

The Early Days of Blotting.- Origins of Protein Blotting.- Western Blotting: Remembrance of Things Past.- Simian Virus 40 and Protein Transfer.- Western Blotting: An Introduction.- From Little Helpers to Automation.- Spectrophotometric Methods to Determine Protein Concentration.- Solubilization of Proteins: The Importance of Lysis Buffer Choice.- Simultaneous Immunoblotting Analysis with Activity Gel Electrophoresis and 2-D Gel Electrophoresis.- Diffusion Blotting: A Rapid and Simple Method for Production of Multiple Blots from a Single Gel.- Multiple Immunoblots by Passive Diffusion of Proteins from a Single SDS-PAGE Gel.- Slice Blotting.- Localizing Proteins by Tissue Printing.- Dot-Immunobinding Assay (Dot-Iba).- Analysis of Antibody Clonotype by Affinity Immunoblotting.- Glycosaminoglycan Blotting and Detection after Electrophoresis Separation.- A Well-Based Reverse-Phase Protein Array of Formalin-Fixed Paraffin-Embedded Tissue.- Quantitative Computerized Western Blotting in Detail.- Cationic Electrophoresis and Eastern Blotting.- A Miniaturized Blotting System for Simultaneous Detecting of Different Autoantibodies.- Proteomic Expressional Profiling of a Paraffin Embedded Tissue by Multiplex Tissue Immunoblotting.- Post-Staining Electroblotting for Efficient and Reliable Peptide Blotting.- Multistrip Western Blotting: A Tool for Comparative Quantitative Analysis of Multiple Proteins.- Western Blotting using PVDF Membranes and Its Downstream Applications.- Blotting from PhastGel to Membranes by Ultrasound.- Western Blotting of High and Low Molecular Weight Proteins Using Heat.- Membrane Strip Affinity Purification of Autoantibodies.- Strip Immunoblotting of Multiple Antigenic Peptides.- Double-Blotting: A Solution to the Problem of Non-Specific Binding of Secondary Antibodies in Immunoblotting Procedures.- Method for Resolution and Western Blotting of Very Large Proteins Using Agarose Electrophoresis.- Immunodetection of P-Selectin Using an Antibody to its C-Terminal Tag.- Improvements and Variants of the Mutliple Antigen Blot Assay-MABA: An Immunoenzymatic Technique for Simultaneous Antigen and Antibody Screening.- Blotting from Immobilized pH Gradient Gels: Application to Total Cell Lysates.- Immunoprecipitation: Western Blot for Proteins of Low Abundance.- Native Electrophoresis and Western Blot Analysis (NEWeB): Methods and Applications.- Shift-Western Blotting: Separate Analysis of Protein and DNA from Protein-DNA Complexes.- Grid-Immunoblotting.- Detection and Quantification of Protein-Protein Interactions by Far-Western Blotting.- Western Blot Analysis of Adhesive Interactions under Fluid Shear Conditions: The Blot Rolling Assay.- Centrifuge Blotting.- Blotting of Coomassie Blue Stained Proteins from PAGE Gels to Transparencies.- B Cell ELISPOT: For the Identification of Antigen-Specific Antibody Secreting Cells.- T Cell ELISPOT: For the Identification of Specific Cytokine-Secreting T Cells.- Membrane Microplates for One- and Two-Color ELISPOT and FLUOROSPOT Assays.- SDS PAGE to Immunoblot in One Hour.- Single Cell Western Blotting.- Protein Detection by Simple Western™ Analysis.- Western Blotting Using Microfluidics.- Two-Dimensional Gel-Based Protein Standardization Verified by Western Blot Analysis.- Fingerprint Deposition of Nitrocellulose and Polyvinylidene Difluoride Membranes Using Alkaline Phosphatase.- Other Notable Protein Blotting Methods: A Brief Review.