This volume compiles a broad range of step-by-step protocols, complementary to the ones published in the first edition of this book, to study various aspects of mitochondrial structure and function in different model organisms, both in vitro and in vivo. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Mitochondria: Practical…mehr
This volume compiles a broad range of step-by-step protocols, complementary to the ones published in the first edition of this book, to study various aspects of mitochondrial structure and function in different model organisms, both in vitro and in vivo. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Mitochondria: Practical Protocols, Second Edition aims to be useful for beginners as well as for experienced researchers in the field.
A Guide to Computational Methods for Predicting Mitochondrial Localization.- Isolation of Functional Mitochondria from Cultured Cells and Mouse Tissues.- Isolation of Mitochondria From Saccharomyces Cerevisiae.- Isolation of Contact Sites Between Inner and Outer Mitochondrial Membranes.- Isolation Of Mitochondria-Associated Membranes (MAM) from Mouse Brain Tissue.- Label-Free Quantitative Analysis of Mitochondrial Proteomes using the Multienzyme Digestion-Filter Aided Sample Preparation (MED-FASP) and 'Total Protein Approach'.- Quantitative Analysis of Glycerophospholipids in Mitochondria by Mass Spectrometry.- Detection of Cysteine Redox States in Mitochondrial Proteins in Intact Mammalian Cells.- Chemical Crosslinking in Intact Mitochondria.- Reconstitution of Mitochondrial Membrane Proteins into Nanodiscs by Cell-Free Expression.- Detection of Dual Targeting and Dual Function of Mitochondrial Proteins in Yeast.- Localizing Mrnas Encoding Mitochondrial Proteins in Yeast byFluorescence Microscopy and Subcellular Fractionation.- Assessing Mitochondrial Bioenergetics in Isolated Mitochondria from Various Mouse Tissues using Seahorse XF96 Analyzer.- Application of FRET-Based Biosensor "Ateam" for Visualization of ATP Levels in the Mitochondrial Matrix of Living Mammalian Cells.- A Microplate-Based Bioluminescence Assay of Mitochondrial Calcium Uptake.- New Imaging Tools to Analyze Mitochondrial Morphology in Caenorhabditis Elegans.- Single Molecule Tracking and Localization Of Mitochondrial Protein Complexes in Live Cells.- Analysis of Yeast Mitochondria by Electron Microscopy.- Analysis of Mitochondrial Membrane Protein Complexes by Electron Cryo-Tomography.- Assays for Mitophagy in Yeast.- Assessing Mitochondrial Selective Autophagy in the Nematode Caenorhabditis Elegans.- Assessing Mitochondrial Unfolded Protein Response in Mammalian Cells.- Analysis of Mitochondrial RNA-Processing Defects in Patient-Derived Tissues by Qrt-PCR and Rnaseq.-Identification of Disease-Causing Mutations by Functional Complementation of Patient-Derived Fibroblast Cell Lines.
A Guide to Computational Methods for Predicting Mitochondrial Localization.- Isolation of Functional Mitochondria from Cultured Cells and Mouse Tissues.- Isolation of Mitochondria From Saccharomyces Cerevisiae.- Isolation of Contact Sites Between Inner and Outer Mitochondrial Membranes.- Isolation Of Mitochondria-Associated Membranes (MAM) from Mouse Brain Tissue.- Label-Free Quantitative Analysis of Mitochondrial Proteomes using the Multienzyme Digestion-Filter Aided Sample Preparation (MED-FASP) and 'Total Protein Approach'.- Quantitative Analysis of Glycerophospholipids in Mitochondria by Mass Spectrometry.- Detection of Cysteine Redox States in Mitochondrial Proteins in Intact Mammalian Cells.- Chemical Crosslinking in Intact Mitochondria.- Reconstitution of Mitochondrial Membrane Proteins into Nanodiscs by Cell-Free Expression.- Detection of Dual Targeting and Dual Function of Mitochondrial Proteins in Yeast.- Localizing Mrnas Encoding Mitochondrial Proteins in Yeast byFluorescence Microscopy and Subcellular Fractionation.- Assessing Mitochondrial Bioenergetics in Isolated Mitochondria from Various Mouse Tissues using Seahorse XF96 Analyzer.- Application of FRET-Based Biosensor "Ateam" for Visualization of ATP Levels in the Mitochondrial Matrix of Living Mammalian Cells.- A Microplate-Based Bioluminescence Assay of Mitochondrial Calcium Uptake.- New Imaging Tools to Analyze Mitochondrial Morphology in Caenorhabditis Elegans.- Single Molecule Tracking and Localization Of Mitochondrial Protein Complexes in Live Cells.- Analysis of Yeast Mitochondria by Electron Microscopy.- Analysis of Mitochondrial Membrane Protein Complexes by Electron Cryo-Tomography.- Assays for Mitophagy in Yeast.- Assessing Mitochondrial Selective Autophagy in the Nematode Caenorhabditis Elegans.- Assessing Mitochondrial Unfolded Protein Response in Mammalian Cells.- Analysis of Mitochondrial RNA-Processing Defects in Patient-Derived Tissues by Qrt-PCR and Rnaseq.-Identification of Disease-Causing Mutations by Functional Complementation of Patient-Derived Fibroblast Cell Lines.
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