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.
About the Author
Table of Contents
1.The Early Days of BlottingEdwin Southern2.Origins of Protein BlottingHarry Towbin3.Western Blotting: Remembrance of Things PastW. Neal Burnette4.Simian Virus 40 and Protein TransferJaime Renart5.Western Blotting: An IntroductionBiji T. Kurien and R. Hal Scofield6.From Little Helpers to AutomationHarry Towbin7.Spectrophotometric Methods to Determine Protein ConcentrationJ.P. Dean Goldring8.Solubilization of Proteins: The Importance of Lysis Buffer ChoiceMandy Peach, Noelle Marsh, Ewa I. Miskiewicz, and Daniel J. MacPhee9.Simultaneous Immunoblotting Analysis with Activity Gel Electrophoresis and 2-D Gel ElectrophoresisDer-Yen Lee and Geen-Dong Chang10.Diffusion Blotting: A Rapid and Simple Method for Production of Multiple Blots from a Single GelIngrid Olsen and Harald G. Wiker11.Multiple Immunoblots by Passive Diffusion of Proteins from a Single SDS-PAGE GelBiji T. Kurien and R. Hal Scofield12.Slice BlottingGraeme Lowe13.Localizing Proteins by Tissue PrintingRafael F. Pont-Lezica14.Dot-Immunobinding Assay (Dot-Iba)Sumi Surendran, Annamma Mathai, and Vishnampet Venkataraman Radhakrishnan15.Analysis of Antibody Clonotype by Affinity ImmunoblottingBiji T. Kurien and R. Hal Scofield16.Glycosaminoglycan Blotting and Detection after Electrophoresis SeparationNicola Volpi and Francesca Maccari17.A Well-Based Reverse-Phase Protein Array of Formalin-Fixed Paraffin-Embedded TissueJoon-Yong Chung and Stephen M. Hewitt18.Quantitative Computerized Western Blotting in DetailDalit Talmi-Frank, Charles L. Jaffe, and Gad Baneth19.Cationic Electrophoresis and Eastern BlottingEngelbert Buxbaum20.A Miniaturized Blotting System for Simultaneous Detecting of Different AutoantibodiesUlrich Canzler, Holger Bartsch, Kai Großmann, Werner Lehmann, Karsten Conrad, Biji T. Kurien, Yaser Dorri, R. Hal Scofield, and Michael P. Bachmann21.Proteomic Expressional Profiling of a Paraffin Embedded Tissue by Multiplex Tissue ImmunoblottingJoon-Yong Chung and Stephen M. Hewitt22.Post-Staining Electroblotting for Efficient and Reliable Peptide BlottingDer-Yen Lee and Geen-Dong Chang23.Multistrip Western Blotting: A Tool for Comparative Quantitative Analysis of Multiple ProteinsEdita Aksamitiene, Jan B. Hoek, and Anatoly Kiyatkin24.Western Blotting using PVDF Membranes and Its Downstream ApplicationsSetsuko Komatsu25.Blotting from PhastGel to Membranes by UltrasoundJoseph Kost and Aharon Azagury26.Western Blotting of High and Low Molecular Weight Proteins Using HeatBiji T. Kuien and R. Hal Scofield27.Membrane Strip Affinity Purification of AutoantibodiesBiji T. Kurien28.Strip Immunoblotting of Multiple Antigenic PeptidesBiji T. Kurien29.Double-Blotting: A Solution to the Problem of Non-Specific Binding of Secondary Antibodies in Immunoblotting ProceduresFrançoise Lasne30.Method for Resolution and Western Blotting of Very Large Proteins Using Agarose ElectrophoresisMarion L. Greaser and Chad M. Warren31.Immunodetection of P-Selectin Using an Antibody to its C-Terminal TagPadmaja Mehta32.Improvements and Variants of the Mutliple Antigen Blot Assay-MABA: An Immunoenzymatic Technique for Simultaneous Antigen and Antibody ScreeningOscar Noya, Sandra Losada, Marilyan Toledo, Adriana Gauna, María Angelita Lorenzo, Henry Bermúdez, and Belkisyolé Alarcón de Noya33.Blotting from Immobilized pH Gradient Gels: Application to Total Cell LysatesHarry Towbin34.Immunoprecipitation: Western Blot for Proteins of Low AbundanceEdward P. Trieu and Ira N. Targoff35.Native Electrophoresis and Western Blot Analysis (NEWeB): Methods and ApplicationsIoannis N. Manoussopoulos and Mina Tsagris36.Shift-Western Blotting: Separate Analysis of Protein and DNA from Protein-DNA ComplexesMatthias Harbers37.Grid-ImmunoblottingOlga Yeliosof and R. Hal Scofield38.Detection and Quantification of Protein-Protein Interactions by Far-Western BlottingJoshua A. Jadwin, Bruce J. Mayer, and Kazuya Machida39.Western Blot Analysis of Adhesive Interactions under Fluid Shear Conditions: The Blot Rolling AssayRobert Sackstein and Robert Fuhlbrigge40.Centrifuge BlottingJinny Paul41.Blotting of Coomassie Blue Stained Proteins from PAGE Gels to TransparenciesJinny Paul42.B Cell ELISPOT: For the Identification of Antigen-Specific Antibody Secreting CellsHemangi B. Shah and Kristi A. Koelsch43.T Cell ELISPOT: For the Identification of Specific Cytokine-Secreting T CellsKerry M. Leehan and Kristi A. Koelsch44.Membrane Microplates for One- and Two-Color ELISPOT and FLUOROSPOT AssaysAlexander E. Kalyuzhny45.SDS PAGE to Immunoblot in One HourBiji T. Kurien, Debashish Danda, Michael Bachmann, and R. Hal Scofield46.Single Cell Western BlottingSyed MS Quadri47.Protein Detection by Simple Western™ AnalysisValerie Harris48.Western Blotting Using MicrofluidicsPothur R. Srinivas49.Two-Dimensional Gel-Based Protein Standardization Verified by Western Blot AnalysisHisao Haniu, Daisuke Watanabe, Yusuke Kawashima, and Hiroyuki Matsumoto50.Fingerprint Deposition of Nitrocellulose and Polyvinylidene Difluoride Membranes Using Alkaline PhosphataseBiji T. Kurien, Debashish Danda, and R. Hal Scofield51.Other Notable Protein Blotting Methods: A Brief ReviewBiji T. Kurien and R. Hal Scofield