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Immunoelectron microscopy is a key technique that bridges the information gap between biochemistry, molecular biology, and ultrastructural studies placing macromolecular functions within a cellular context. In Immunoelectron Microscopy: Methods and Protocols, expert researchers combine the tools of the molecular biologist with those of the microscopist. From the molecular biology toolbox, this volume presents methods for antigen production by protein expression in bacterial cells, methods for epitope tagged protein expression in plant and animal cells allowing protein localization in the absence of protein specific antibodies as well as methods for the production of anti-peptide, monoclonal, and polyclonal antibodies. From the microscopy toolbox, sample preparation methods for cells, plant, and animal tissue are presented. Both cryo-methods, which have the advantage of retaining protein antigenicity at the expense of ultrastructural integrity, as well as chemical fixation methods that maintain structural integrity while sacrificing protein antigenicity have been included, with chapters examining various aspects of immunogold labeling. Written in the highly successful Methods in Molecular Biology{u0CE5}ries format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and essential, Immunoelectron Microscopy: Methods and Protocols seeks to facilitate an increased understanding of structure function relationships. Presents a wide range of Cryo and chemical fixation methods for single cells, plant, and animal tissue Contains Immunogold labeling methods for transmission and scanning electron microscopy Includes comprehensive pre and post embedding immunolabeling methods

Índice: Protein Antigen Expression in E. coli for Antibody Production.- Expression of Epitope-Tagged Proteins in Plants.- Expression of Epitope-Tagged Proteins in Arabidopsis Leaf Mesophyll Protoplasts.- Transient Expression of Epitope-Tagged Proteins in Mammalian Cells.- Production and Purification of Polyclonal Antibodies.- Production and Purification of Monoclonal Antibodies.- Production of Antipetide Antibodies.- Preparation of Colloidal Gold Particles and Conjugation to Protein A, IgG, F(ab)̕2 and Strepavidin.- Immunoelectron Microscopy of Chemically Fixed Developing Plant Embryos.- Pre-Embedding Immunogold L... Etc.