Rab GTPases Methods and Protocols

This second edition volume expands on the previous edition with a discussion of new research and discoveries in the Rab field. Chapters in this book cover topics such as new information on Rab regulation and localization; interaction; function; and diseases. 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. 

Cutting-edge and comprehensive, Rab GTPases: Methods and Protocols, Second Edition is a valuable resource for scientists working in the fields of Rab and other small GTPases, and beyond.

Newer Methods Drive Recent Insights into Rab GTPase Biology: An Overview.- Rab29 Fast Exchange Mutants: Characterization of a Challenging Rab GTPase.- High-Throughput Assay for Profiling the Substrate Specificity of Rab GTPase-Activating Proteins.- Detecting Endogenous Rab8 Activation.- Testing the Phenotypic Effects of a Rab Chimera that Resolves Exchange Factor Specificity from Effector Specificity.- Profiling Structural Alterations during Rab5 Nucleotide Exchange by HDX-MS.- CLEM Characterization of Rab8 and Associated Membrane Trafficking Regulators at Primary Cilium Structures.- Imaging of Spatial Cycling of Rab GTPase in the Cell.- Deconvolution of Multiple Rab Binding Domains using the Batch Yeast 2-Hybrid Method DEEPN.- Determination of the Rab27-Effector Binding Affinity Using a High-Throughput FRET-Based Assay.- Methods to Study the Unique SOCS Box Domain of the Rab40 Small GTPase Subfamily.- Using GBP Nanotrap to Restore Autophagy in the Rab5/Vps21 

Mutant by Forcing Snf7 and Atg17 Interaction.- Establishing Regulation of a Dynamic Process by Ypt/Rab GTPase: A Case for Cisternal Progression.- Methods for Assessing the Regulation of a Kinase by the Rab GTPase Ypt1.- Qualitative and Quantitative Assessment of the Role of Endocytic Regulatory and/or Rab Proteins in Mitochondrial Fusion and Fission.- Characterization of the Role of Rab18 in Mediating LD-ER Contact and LD Growth.- Methods for Establishing Rab Knockout MDCK Cells.- Generating Rab6 Conditional Knockout Mice.- Use of Immunohistochemistry to Determine Expression of Rab5 Subfamily 

of GTPases in Mature and Developmental Brains.- Assessing Rab5 Activation in Health and Disease.- Quantitative Fluorescence Microscopy for Detecting Mammalian Rab Vesicles within the Parasitophorous Vacuole of the HumanPathogen Toxoplasma gondii.