Cytokine Bioassays pp 173-184 | Cite as
A Mix-and-Measure Assay for Determining the Activation Status of Endogenous Cdc42 in Cytokine-stimulated Macrophage Cell Lysates
Protocol
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Abstract
Cytokine stimulations of leukocytes many times result in transient activation of the p21 Rho family of small GTPases. The role of these molecules during cell migration and chemotaxis is well established. The traditional approach to study the activation dynamics of these proteins involves affinity pull-downs that are often cumbersome and prone to errors. Here, we describe a reagent and a method of simple “mix-and-measure” approach useful for determining the activation status of endogenous Cdc42 GTPase from cell lysates.
Key words
Cdc42 activity Biosensor MeroCBD Fluorometry Mix-and-measureNotes
Acknowledgement
This work was supported by grants GM093121 (D.S., L.H.), T32GM007491 (V.M.), and GM071828 (D.C.).
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© Springer Science+Business Media, New York 2014