A mix-and-measure assay for determining the activation status of endogenous Cdc42 in cytokine-stimulated macrophage cell lysates

Veronika Miskolci, Désirée Spiering, Dianne Cox, Louis Hodgson

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)173-184
Number of pages12
JournalMethods in Molecular Biology
Volume1172
DOIs
StatePublished - 2014
Externally publishedYes

Keywords

  • Biosensor
  • Cdc42 activity
  • Fluorometry
  • MeroCBD
  • Mix-and-measure

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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