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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 (TM) series 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.
This volume presents authoritative and cutting-edge methods and protocols focusing on three tool boxes covering the increasingly diverse methodologies used to image selected proteins and to investigate their function by light and electron microscopy. The first tool box includes the development of a wide range of molecular and immunological probes to target specific proteins. The second details the use of these probes for high resolution fluorescence microscopy and the third focuses on applications for transmission and scanning electron microscopy. 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 thus ensuring successful results in the further study of this vital field.
This much-needed book is the first definitive volume on Euglena in twenty-fire years, offering information on its atypical biochemistry, cell and molecular biology, and potential biotechnology applications. This volume gathers together contributions from well-known experts, who in many cases played major roles in elucidating the phenomenon discussed. Presented in three parts, the first section of this comprehensive book describes novel biochemical pathways which in some instances have an atypical subcellular localization. The second section details atypical cellular mechanisms of organelle protein import, organelle nuclear genome interdependence, gene regulation and expression that provides insights into the evolutionary origins of eukaryotic cells. The final section discusses how biotechnologists have capitalized on the novel cellular and biochemical features of Euglena to produce value added products. Euglena: Biochemistry, Cell and Molecular Biology will provide essential reading for cell and molecular biologists with interests in evolution, novel biochemical pathways, organelle biogenesis and algal biotechnology. Readers will come away from this volume with a full understanding of the complexities of the Euglena as well as new realizations regarding the diversity of cellular processes yet to be discovered.
This much-needed book is the first definitive volume on Euglena in twenty-fire years, offering information on its atypical biochemistry, cell and molecular biology, and potential biotechnology applications. This volume gathers together contributions from well-known experts, who in many cases played major roles in elucidating the phenomenon discussed. Presented in three parts, the first section of this comprehensive book describes novel biochemical pathways which in some instances have an atypical subcellular localization. The second section details atypical cellular mechanisms of organelle protein import, organelle nuclear genome interdependence, gene regulation and expression that provides insights into the evolutionary origins of eukaryotic cells. The final section discusses how biotechnologists have capitalized on the novel cellular and biochemical features of Euglena to produce value added products. Euglena: Biochemistry, Cell and Molecular Biology will provide essential reading for cell and molecular biologists with interests in evolution, novel biochemical pathways, organelle biogenesis and algal biotechnology. Readers will come away from this volume with a full understanding of the complexities of the Euglena as well as new realizations regarding the diversity of cellular processes yet to be discovered.
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 (TM) series 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.
This volume presents authoritative and cutting-edge methods and protocols focusing on three tool boxes covering the increasingly diverse methodologies used to image selected proteins and to investigate their function by light and electron microscopy. The first tool box includes the development of a wide range of molecular and immunological probes to target specific proteins. The second details the use of these probes for high resolution fluorescence microscopy and the third focuses on applications for transmission and scanning electron microscopy. 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 thus ensuring successful results in the further study of this vital field.
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