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This volume presents a range of different techniques that have been used to characterize the structure and function of the endoplasmic reticulum (ER) in higher plants. Chapters guide readers through application of modern microscopy techniques by fluorescence and electron microscopy, new protocols for analysing ER network structure, methods to purify and analyse ER membrane structure and to study protein glycosylation, protocols to study the unfolded protein response, and the role of the ER in autophagy. 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.
Authoritative and cutting-edge, The Plant Endoplasmic Reticulum: Methods and Protocols aims to ensure successful results in the further study of this vital field.
List of contents
Labelling the ER for Light and Fluorescence Microscopy.- 3D Electron Microscopy of the ER.- Characterization of Proteins Localised to Plant ER-PM Contact Sites.- Preparation and Imaging of Specialised ER using Super-resolution and TEM Techniques.- Quantitation of ER Structure and Function.- Long-term Imaging of Endoplasmic Reticulum Morphology in Embryos during Seed Germination .- Dancing with the Stars: Using Image Analysis to Study the Choreography of the Endoplasmic Reticulum and Its Partners and of Movement within its Tubules.- Preparation of Highly Enriched ER Membranes using Free-flow Electrophoresis.- ER Microsome Preparation in Arabidopsis thaliana.- ER Membrane Lipid Composition and Metabolism: Lipidomic Analysis.- 2in1 Vectors Improve in Planta BiFC and FRET Analysis.- Metabolons on the Plant ER.- Using Optical Tweezers Combined with Total Internal Reflection Microscopy to Study Interactions between the ER and Golgi in Plant Cells.- Protein Biosynthesis and Maturationin the ER.- ER Membrane Protein Interactions using the Split-Ubiquitin System (SUS).- Analysis of Protein Glycosylation in the ER.- The Unfolded Protein Response.- Unfolded Protein Response in Arabidopsis.- Fluorescence Imaging of Autophagy-mediated Er-to-vacuole Trafficking in Plants.- Imaging the ER and Endomembrane System in Cereal Endosperm.
Summary
This volume presents a range of different techniques that have been used to characterize the structure and function of the endoplasmic reticulum (ER) in higher plants. Chapters guide readers through application of modern microscopy techniques by fluorescence and electron microscopy, new protocols for analysing ER network structure, methods to purify and analyse ER membrane structure and to study protein glycosylation, protocols to study the unfolded protein response, and the role of the ER in autophagy. 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.
Authoritative and cutting-edge, The Plant Endoplasmic Reticulum: Methods and Protocols aims to ensure successful results in the further study of this vital field.
