title: A mix-and-measure assay for determining the activation status of endogenous Cdc42 in cytokine-stimulated macrophage cell lysates.
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.
authors.affiliation: Department of Anatomy and Structural Biology, Albert Einstein College of Medicine of Yeshiva University, 1300 Morris Park Ave, Bronx, NY, 10461, USA, veronika.miskolci@phd.einstein.yu.edu.
authors.forename: Veronika
authors.initials: V
authors.lastname: Miskolci
authors.forename: Désirée
authors.initials: D
authors.lastname: Spiering
authors.forename: Dianne
authors.initials: D
authors.lastname: Cox
authors.forename: Louis
authors.initials: L
authors.lastname: Hodgson
mesh_terms: D000818:Animals; D001681:Biological Assay; D002457:Cell Extracts / Q000737:chemistry; D002460:Cell Line; D054428:Chemokine CX3CL1 / Q000494:pharmacology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008262:Macrophage Activation / Q000187:drug effects*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*; D020764:cdc42 GTP-Binding Protein / Q000235:genetics* / Q000378:metabolism
chemical_list: C581598:Cdc42 protein, mouse; D002457:Cell Extracts; D054428:Chemokine CX3CL1; C515539:Cx3cl1 protein, mouse; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D020764:cdc42 GTP-Binding Protein