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Focusing on new methods and techniques developed to address longstanding questions about the Golgi complex, this volume explores a diverse set of chapters, ranging from live and fixed cell imaging techniques toin vitro biochemical reconstitution systems. Each chapter provides a detailed set of specific instructions, which should enable anyone to successfully complete the assays. Written for the highly successful Methods in Molecular Biology series program, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily…mehr
Focusing on new methods and techniques developed to address longstanding questions about the Golgi complex, this volume explores a diverse set of chapters, ranging from live and fixed cell imaging techniques toin vitro biochemical reconstitution systems. Each chapter provides a detailed set of specific instructions, which should enable anyone to successfully complete the assays. Written for the highly successful Methods in Molecular Biology series program, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips from the experts on troubleshooting and avoiding known pitfalls.
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Autorenporträt
WILLIAM J. BROWN works for Saga Software Inc. in Reston, Virginia, as Director of Research andDevelopment for the Sagavista enterprise integration broker.
Inhaltsangabe
4D Confocal Imaging of Yeast Organelles.- Imaging the Polarized Sorting of Proteins from the Golgi Complex in Live Neurons.- Imaging Golgi Outposts in Fixed and Living Neurons.- Analysis of Arf1 GTPase-Dependent Membrane Binding and Remodeling Using the Exomer Secretory Vesicle Cargo Adaptor.- STEM Tomography Imaging of Hypertrophied Golgi Stacks in Mucilage-Secreting Cells.- Reconstitution of COPI Vesicle and Tubule Formation.- Reconstitution of Phospholipase A2-Dependent Golgi Membrane Tubules.- Proteomic Characterization of Golgi Membranes Enriched from Arabidopsis Suspension Cell Cultures.- High Content Analysis of the Golgi Complex by Correlative Screening Microscopy.- Activity Detection of GalNAc Transferases by Protein-Based Fluorescence Sensors In Vivo.- In Situ Proximity Ligation Assay (PLA) Analysis of Protein Complexes Formed Between Golgi-Resident, Glycosylation-Related Transporters and Transferases in AdherentMammalian Cell Cultures.- Creating Knock-Outs of Conserved Oligomeric Golgi Complex Subunits Using CRISPR-Mediated Gene Editing Paired with a Selection Strategy Based on Glycosylation Defects Associated with Impaired COG Complex Function.- Reversible Controlled Aggregation of Golgi Resident Enzymes to Assess Their Transport/Dynamics Along the Secretory Pathway.- Assays to Study the Fragmentation of the Golgi Complex During the G2/M Transition of the Cell Cycle.- The Role of Lysophospholipid Acyltransferases in the Golgi Complex.- Methods to Purify and Assay Secretory Pathway Kinases.
4D Confocal Imaging of Yeast Organelles.- Imaging the Polarized Sorting of Proteins from the Golgi Complex in Live Neurons.- Imaging Golgi Outposts in Fixed and Living Neurons.- Analysis of Arf1 GTPase-Dependent Membrane Binding and Remodeling Using the Exomer Secretory Vesicle Cargo Adaptor.- STEM Tomography Imaging of Hypertrophied Golgi Stacks in Mucilage-Secreting Cells.- Reconstitution of COPI Vesicle and Tubule Formation.- Reconstitution of Phospholipase A2-Dependent Golgi Membrane Tubules.- Proteomic Characterization of Golgi Membranes Enriched from Arabidopsis Suspension Cell Cultures.- High Content Analysis of the Golgi Complex by Correlative Screening Microscopy.- Activity Detection of GalNAc Transferases by Protein-Based Fluorescence Sensors In Vivo.- In Situ Proximity Ligation Assay (PLA) Analysis of Protein Complexes Formed Between Golgi-Resident, Glycosylation-Related Transporters and Transferases in AdherentMammalian Cell Cultures.- Creating Knock-Outs of Conserved Oligomeric Golgi Complex Subunits Using CRISPR-Mediated Gene Editing Paired with a Selection Strategy Based on Glycosylation Defects Associated with Impaired COG Complex Function.- Reversible Controlled Aggregation of Golgi Resident Enzymes to Assess Their Transport/Dynamics Along the Secretory Pathway.- Assays to Study the Fragmentation of the Golgi Complex During the G2/M Transition of the Cell Cycle.- The Role of Lysophospholipid Acyltransferases in the Golgi Complex.- Methods to Purify and Assay Secretory Pathway Kinases.
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