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Rapid parallel flow cytometry assays of active GTPases using effector beads.


ABSTRACT: We describe a rapid assay for measuring the cellular activity of small guanine triphosphatases (GTPases) in response to a specific stimulus. Effector-functionalized beads are used to quantify in parallel multiple GTP-bound GTPases in the same cell lysate by flow cytometry. In a biologically relevant example, five different Ras family GTPases are shown for the first time to be involved in a concerted signaling cascade downstream of receptor ligation by Sin Nombre hantavirus.

SUBMITTER: Buranda T 

PROVIDER: S-EPMC3848308 | biostudies-other | 2013 Nov

REPOSITORIES: biostudies-other

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Rapid parallel flow cytometry assays of active GTPases using effector beads.

Buranda Tione T   BasuRay Soumik S   Swanson Scarlett S   Agola Jacob J   Bondu Virginie V   Wandinger-Ness Angela A  

Analytical biochemistry 20130806 2


We describe a rapid assay for measuring the cellular activity of small guanine triphosphatases (GTPases) in response to a specific stimulus. Effector-functionalized beads are used to quantify in parallel multiple GTP-bound GTPases in the same cell lysate by flow cytometry. In a biologically relevant example, five different Ras family GTPases are shown for the first time to be involved in a concerted signaling cascade downstream of receptor ligation by Sin Nombre hantavirus. ...[more]

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