Immunofluorescent detection of the activation of the small GTPase Rac1 in mouse skeletal muscle fibers

Nobuyuki Takenaka; Yuma Nihata; Takaya Satoh

doi:10.1016/j.ab.2014.09.013

Immunofluorescent detection of the activation of the small GTPase Rac1 in mouse skeletal muscle fibers

Anal Biochem. 2015 May 1:476:5-7. doi: 10.1016/j.ab.2014.09.013. Epub 2014 Sep 30.

Authors

Nobuyuki Takenaka¹, Yuma Nihata¹, Takaya Satoh²

Affiliations

¹ Laboratory of Cell Biology, Department of Biological Science, Graduate School of Science, Osaka Prefecture University, Naka-ku, Sakai 599-8531, Japan.
² Laboratory of Cell Biology, Department of Biological Science, Graduate School of Science, Osaka Prefecture University, Naka-ku, Sakai 599-8531, Japan. Electronic address: tkysato@b.s.osakafu-u.ac.jp.

PMID: 25277816
DOI: 10.1016/j.ab.2014.09.013

Abstract

The small GTPase Rac1 acts as a molecular switch of intracellular signaling in mammals. For understanding the regulatory mechanism, it is important to identify subcellular locations in which Rac1 is activated following multiple extracellular stimuli. However, it is difficult to detect Rac1 activation in situ in animal tissues, and thus a novel method is highly desirable. Here, we report a simple method to visualize the activation of endogenous Rac1 in mouse skeletal muscle fibers. In this assay, specific interaction between activated Rac1 and a binding polypeptide is detected by immunofluorescent microscopy. This approach is readily applicable to other small GTPases.

Keywords: Activated form; Immunofluorescent microscopy; Rac1; Skeletal muscle; Small GTPase; Subcellular localization.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Humans
In Vitro Techniques
Male
Mice
Mice, Inbred C57BL
Microscopy, Fluorescence
Monomeric GTP-Binding Proteins / metabolism*
Muscle, Skeletal / enzymology*
rac1 GTP-Binding Protein / metabolism

Substances

Monomeric GTP-Binding Proteins
rac1 GTP-Binding Protein