Peroxisomes: Methods and Protocols

by Michael Schrader (Editor)

Overview

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.

Product Details

ISBN-13:	9781493983445
Publisher:	Springer New York
Publication date:	07/20/2018
Series:	Methods in Molecular Biology , #1595
Edition description:	Softcover reprint of the original 1st ed. 2017
Pages:	347
Product dimensions:	7.01(w) x 10.00(h) x (d)

Table of Contents

Isolation of Mammalian Peroxisomes by Density Gradient Centrifugation.- Analysis of Yeast Peroxisomes via Spatial Proteomics.- Isolation of Glycosomes from Trypanosoma brucei.- Immunolabeling for Detection of Endogenous and Overexpressed Peroxisomal Proteins in Mammalian Cells.- Super-Resolution Imaging of Peroxisomal Proteins Using STED Nanoscopy.- Direct Shastic Optical Reconstruction Microscopy (dSTORM) of Peroxisomes.- Correlative Light- and Electron Microscopy in Peroxisome Research.- Ultrastructural Analysis and Quantification of Peroxisome-Organelle Contacts.- Detection of Peroxisomal Proteins during Mycobacterial Infection.- Proximity-Ligation Assay to Detect Peroxisome-Organelle Interaction.- Assay of Reactive Oxygen/Nitrogen Species (ROS/RNS) in Arabidopsis Peroxisomes through Fluorescent Protein Containing a Type 1-Peroxisomal Targeting Signal (PTS1).- Identification of Peroxisome-Derived Hydrogen Peroxide-Sensitive Target Proteins Using a YAP1C-Based GeneticProbe.- Assessment of the Peroxisomal Redox State in Living Cells Using NADPH- and NAD⁺/NADH-Specific Fluorescent Protein Sensors.- Live-Cell Imaging of Peroxisomal Calcium Levels and Dynamics.- Analysis of Peroxisome Biogenesis by Phos-Tag SDS-PAGE.- Targeted Modifications of the Yeast Genome to Study Peroxisomes.- Applying CRISPR-Cas9 Genome Editing to Study Genes Involved in Peroxisome Biogenesis or Peroxisomal Functions.- Generation of Reporter Cell Lines for Endogenous Expression Analysis of Peroxisomal Proteins.- Peroxisomes and Viruses: Overview on Current Knowledge and Experimental Approaches.- Tools to Investigate the Peroxisome-Dependent Antiviral Response.- Determining the Importance of Peroxisomal Proteins for Viral Infections in Cultured Mammalian Cells.- Characterization of Yeast Peroxisomes: Enrichment of Peroxisomal Fractions and Analysis of b-Oxidation Activity.- A Cell-Free In Vitro Import System for Peroxisomal Proteins Containing a Type 2 Targeting Signal (PTS2).- Assessing Peroxisomal Protein Interaction by Immunoprecipitation.- Purification of a Recombinant Human PEX1/PEX6 AAA+ ATPase Complex from HEK293TT Cells.- Affinity Purification of Soluble and Membrane-Bound Protein Complexes by a FlpIn Strategy.- Utilization of Nonstop mRNA to Assess Ribosome-Associated Nascent Polypeptide Chains in Early Topogenesis of Peroxisomal Proteins.- Computational Evaluation of Peroxisomal Targeting Signals in Metazoa.- Computational Approaches for Peroxisomal Protein Localization.- Estimating the Interaction Strength between PTS1-Peptides and Their Receptor PEX5 in Living Cells Using Flow-Cytometer-Based FRET (flowFRET) Measurements.- Using the Super Folder GFP (sfGFP) System to Study Plant Peroxisomal Protein Import.- Biochemical Fractionation of Trypanosomes for the Analysis of Glycosomal Protein Import Defects.- Manipulation and Visualization of Peroxisomes in Drosophila.- Mouse Models to Study Peroxisomal Functions and Disorders: Overview,Caveats, and Recommendations.