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This second edition offers 88 chapters divided among three volumes providing the most comprehensive source of know-how in the wide-ranging field of Mitochondrial Medicine. Volume II guides readers through chapters on mitochondrial dysfunction, functional' mitochondria, mitochondrial retrograde, mitochondrial dNTP pool quantification, mitochondrial ADP-ribosylation, blue-native gel approach, 3D optical cryo-imaging method, mitochondrial ATP and ROS production, protocol for untargeted metabolomic analysis, and methods for analysis of nitrotyrosine-containing proteins. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and accessible, Mitochondrial Medicine, Second Edition, Volume 2: Assessing Mitochondria aims to be acomprehensive source of know-how in the wide-ranging field of Mitochondrial Medicine.
List of contents
Mitochondrial Dysfunction in Mitochondrial Medicine: Current Limitations, Pitfalls, and Tomorrow.- Preparation of 'Functional' Mitochondria - A Challenging Business.- Isolation and Quality Control of Functional Mitochondria.- Purification of Functional Platelet Mitochondria Using a Discontinuous Percoll Gradient.- Mechanical Permeabilization as a New Method for Assessment of Mitochondrial Function in Insect Tissues.- Analysis of Mitochondrial Retrograde Signaling In Yeast Model Systems.- Native Gel Electrophoresis and Immunoblotting to Analyze Electron Transport Chain Complexes.- Measuring Mitochondrial Hydrogen Peroxide Levels and Glutathione Redox Equilibrium in Drosophila Neuron Subtypes Using Redox-Sensitive Fluorophores and 3D Imaging.- Assessment of Mitochondrial Cell Metabolism By Respiratory Chain Electron Flow Assays.- Whole-Cell and Mitochondrial dNTP Pool Quantification from Cells and Tissues.- Single Particle-Tracking Method in Fluorescence Microscopy to Monitor Bioenergetic Responses Of Individual Mitochondria.- Investigation of Mitochondrial ADP-ribosylation via Immunofluorescence.- Assessment of Mitochondrial Ca2+ Uptake.- Assessment of Mitochondrial Membrane Potential and NADH Redox State in Acute Brain Slices.- Evaluation of Mitochondria Content and Function in Live Cells by Multi-color Flow Cytometric Analysis.- Analysis of Mitochondrial Dysfunction during Cell Death.- Modified Blue-Native Gel Approach for Analysis of Respiratory Super-Complexes.- Patch-Clamp Recording of the Activity of Ion Channels in the Inner Mitochondrial Membrane.- Assessment of Mitochondrial Protein Glutathionylation as Signaling for CO Pathway.- 3D Optical Cryo-Imaging Method: A Novel Approach To Quantify Renal Mitochondrial Bioenergetics Dysfunction.- Simultaneous Quantification of Mitochondrial ATP and ROS Production Using ATP Energy Clamp Methodology.- High-Throughput Image Analysis of Lipid-Droplet-Bound Mitochondria.- Cell Energy Budget Platform for Multi-Parametric Assessment of Cell and Tissue Metabolism.- Fluorescence-based Assay for Measuring OMA1 Activity.- Studying Mitochondrial Network Formation by in vivo and in vitro Reconstitution Assay.- Extraction of Functional Mitochondria Based on the Membrane Stiffness.- A Protocol for Untargeted Metabolomic Analysis: From Sample Preparation to Data Processing.- A Method for Analysis of Nitrotyrosine-Containing Proteins by Immunoblotting Coupled With Mass Spectrometry.- In vivo Visualization And Quantification of Mitochondrial Morphology in C. elegans.- Assessing Impact of Platinum Complexes on Mitochondrial Functions.- In-silico Modeling of the Mitochondrial Pumping Complexes with Markov State Models.- Monitoring the Mitochondrial Presequence Import Pathway in Living Mammalian Cells with a NewMolecular Biosensor.