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This volume provides easily accessible and comprehensive collection of methods, techniques, and strategies to investigate the molecular and cellular biology of peroxisomes in different organisms. Chapters detail valuable instructions, guidelines and protocols for molecular cell biologists, biochemists and biomedical researchers with an interest in peroxisome biology. Chapters in Peroxisomes: Methods and Protocols illustrate the isolation of peroxisomes, investigation of properties of membrane proteins, protocols to investigate and manipulate peroxisomes in cellular systems, detection of peroxisomes, including immunofluorescence, cytochemistry, cryo-immuno electron microscopy, and live cell imaging approaches.
Authoritative and practical, Peroxisomes: Methods and Protocols aims to be useful for those already working on peroxisomes as well as for those who would like to start working on this fascinating organelle.
List of contents
Isolation of Peroxisomes from Rat Liver and Cultured Hepatoma Cells by Density Gradient Centrifugation.- Isolation of Peroxisomes from Mouse Brain Using a Continuous Nycodenz Gradient: A Comparison to the Isolation of Liver and Kidney Peroxisomes.- Determining the Topology of Peroxisomal Proteins Using Protease Protection Assays.- Isolation of Native Soluble and Membrane-Bound Protein Complexes from Yeast Saccharomyces Cerevisiae.- Method for Measurement of Peroxisomal Very Long-Chain Fatty Acid Beta-Oxidation and de novo C26:0 Synthesis Activity in Living Cells Using Stable-Isotope Labelled Docosanoic Acid.- Analysis of Plasmalogen Synthesis in Cultured Cells.- Transfection of Primary Human Skin Fibroblasts for Peroxisomal Studies.- siRNA-Mediated Silencing of Peroxisomal Genes in Mammalian Cells.- Dual Reporter Systems for the Analysis of Translational Readthrough in Mammals.- Cytochemical Detection of Peroxisomes in Light and Electron Microscopy with 3,3'-diaminobenzidine.- Cryo-Immuno Electron Microscopy of Peroxisomal Marker Proteins.- Detection and Immunolabelling of Peroxisomal Proteins.- Labelling of Peroxisomes for Live Cell Imaging in the Filamentous Fungus Ustilago Maydis.- Quantitative Monitoring of Subcellular Redox Dynamics in Living Mammalian Cells Using RoGFP2-based Probes.- KillerRed as a Tool to Study the Cellular Responses to Peroxisome-Derived Oxidative Stress.- Determination of Peroxisomal pH in Living Mammalian Cells Using pHRed.- In Cellulo Approaches to Study Peroxisomal Protein Import - Yeast Immunofluorescence Microscopy.- Blue Native Page: Applications to Study Peroxisome Biogenesis.- In Vitro PMP Import Analysis Using Cell-Free Synthesized PMP and Isolated Peroxisomes.- Peroxisomal Membrane and Matrix Protein Import Using a Semi-Intact Mammalian Cell System.- The Use of Glycosylation Tags as Reporters for Protein Entry into the Endoplasmic Reticulum in Yeast and Mammalian Cells.- Detection of Ubiquitinated Peroxisomal Proteins in Yeast.- Assessing Pexophagy in Mammalian Cells.- Experimental Systems to Study Yeast Pexophagy.- Flow Cytometric Analysis of the Expression Pattern of Peroxisomal Proteins, Abcd1, Abcd2, and Abcd3 in BV-2 Murine Microglial Cells.- Study of Peroxisomal Protein Phosphorylation by Functional Proteomics.- Analysis of Peroxisomal beta-Oxidation during Storage Oil Mobilization in Arabidopsis Thaliana Seedlings.- Peroxisome Mini-Libraries - Systematic Approaches to Study Peroxisomes Made Easy.- Generation of Peroxisome-Deficient Somatic Animal Cell Mutants.- Clinical and Laboratory Diagnosis of Peroxisomal Disorders.
Summary
This volume provides easily accessible and comprehensive collection of methods, techniques, and strategies to investigate the molecular and cellular biology of peroxisomes in different organisms. Chapters detail valuable instructions, guidelines and protocols for molecular cell biologists, biochemists and biomedical researchers with an interest in peroxisome biology. Chapters in Peroxisomes: Methods and Protocols illustrate the isolation of peroxisomes, investigation of properties of membrane proteins, protocols to investigate and manipulate peroxisomes in cellular systems, detection of peroxisomes, including immunofluorescence, cytochemistry, cryo-immuno electron microscopy, and live cell imaging approaches.
Authoritative and practical, Peroxisomes: Methods and Protocols aims to be useful for those already working on peroxisomes as well as for those who would like to start working on this fascinating organelle.
