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Full Description
This detailed book expands upon the previous edition with a collection of methods for those performing experimental work on small GTPases of the Rho family. Split into four sections, the volume explores computational modeling and imaging procedures, biochemical methods related to post-translational modifications of Rho GTPases as well as some high throughput methods, functional assays that allow for monitoring the consequences of manipulating Rho GTPases in a variety of cell types and cell biology processes, and techniques specifically designed for studies in selected non-mammalian model organisms (zebrafish, social ameba, plants and algae). Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on trouble shooting and avoiding known pitfalls.
Authoritative and up-to-date, Rho GTPases: Methods and Protocols, Second Edition constitutes an invaluable tool for all those with an interest in this remarkable family of signaling proteins.
Contents
Computational Modeling of the Dynamics of Spatiotemporal Rho GTPase Signaling: A Systematic Review.- Uncovering Bistability in the Rac1/RhoA Signaling Network through Integrating Computational Modeling and Experimentation.- Modeling Rho GTPase Dynamics Using Boolean Logic.- EdgeProps: A Computational Platform for Correlative Analysis of Cell Dynamics and Near-Edge Protein Activity.- Applying Perturbation Expectation-Maximization to Protein Trajectories of Rho GTPases.- Parallel High Resolution Imaging of Leukocyte Chemotaxis under Agarose with Rho Family GTPase Biosensors.- Characterization of Genetically-Encoded FRET Biosensors for Rho Family GTPases.- Screening for Rho GTPase Modulators in Actin-Dependent Processes Exemplified by Phagocytosis.- An In Vitro Kinase Assay to Assess Rac1 Phosphorylation by ERK.- Methods to Study Rho GTPases Using Bacterial Toxins.- Assessment of S-Glutathionylated Rac1 in Cells Using Biotin-Labeled Glutathione.- High Throughput Assay for RhoGEFs Based on the Transcreener® GDP Assay.- Small Volume Flow Cytometry-Based Multiplex Analysis of the Activity of Small GTPases.- Methods to Study the Roles of Rho GTPases in Platelet Function.- Methods to Investigate the Role of Rho GTPases in Osteoclast Function.- Methods to Study the Role of Cdc42, Rac1 and Rac2 in B Cell Cytoskeletal Responses.- In Vivo Rescue Assay of RhoA Deficient Hematopoietic Stem and Progenitor Cells.- Cell Polarity and Division Symmetry Analyses in Transformed Blood Cells.- C3 Transferase Gene Therapy for Continuous RhoA Inhibition.- A Practical Protocol for the Conditional Depletion of Rho Isoforms in Human Embryonic Stem Cells.- Methods to Study the Roles of Rho GTPases in Dendritic Tree Complexity.- Assessing the Roles of Rho GTPases in Cell DNA Repair by the Nucleotide Excision Repair Pathway.- Measuring the Contributions of the Rho Pathway to the DNA Damage Response in Tumor Epithelial Cells.- Using GAL4-Inducible Transgenics to Modulate Rho GTPase Activity in Zebrafish.- Assaying Rho GTPase-Dependent Processes in Dictyostelium discoideum.- Reconstruction of ROP GTPase Domains on the Plasma Membrane in Tobacco Leaves.- Analyzing Rho GTPase-Dependent Processes during Cell Polarization in Brown Algae.
