Titlebook: Rho GTPases; Methods and Protocol Francisco Rivero Book 2018Latest edition Springer Science+Business Media, LLC, part of Springer Nature 20

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Methods to Study the Role of Cdc42, Rac1, and Rac2 in B-Cell Cytoskeletal Responses adhesion. Spreading of B cells is dependent on Cdc42, while Rac1 and Rac2 are necessary for homotypic adhesion. These in vitro assays can be used to examine functional responses of B cells mediated by the cell cytoskeleton, for example when comparing B cells from different gene knockout animals.

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Modeling Rho GTPase Dynamics Using Boolean Logic the network into appropriate simulation software (CellNetAnalyzer); and finally perform simulations and make novel, testable predictions via in silico knockouts. Given this predictive power, the Boolean approach may ultimately help to highlight potential future avenues of experimental research.

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An In Vitro Kinase Assay to Assess Rac1 Phosphorylation by ERKactivity of the kinase EKR. This in vitro kinase assay uses commercially available purified active ERK. Substrate Rac1 was generated and purified as a glutathione S-transferase (GST) fusion protein. [γ-.P]ATP is used to radiolabel Rac1. Phosphorylation of Rac1 is viewed by autoradiography.

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