Kurien / Scofield Western Blotting

Methods and Protocols
2015
ISBN: 978-1-4939-2694-7
Verlag: Springer US
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 1312, 509 Seiten, eBook

Reihe: Methods in Molecular Biology

This volume covers past and present western blot techniques, such as diffusion blotting, slice blotting, blotting of high and low molecular weight proteins, single cell blotting and automated blotting. 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. Thorough and cutting-edge, Western Blotting: Methods and Protocols will serve as an invaluable reference for those interested in further study into this fascinating field.

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Kurien, Biji T.

Scofield, R. Hal

Inhaltsverzeichnis

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.

Über Autor(innen)

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: [phone] ext 35204 Email: [e-mail] 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: [phone] Fax: [phone] E-mail: [e-mail]