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Sci. STKE, 3 June 2003
Vol. 2003, Issue 185, p. pl10
[DOI: 10.1126/stke.2003.185.pl10]

PROTOCOLS

Analysis of the Mobility of Signaling Molecules in Lymphocytes Using Fluorescence Photobleaching Techniques

Natsuko Tanimura1, Masakazu Nagafuku1, Douglas R. Liddicoat1, Toshiyuki Hamaoka2, and Atsushi Kosugi1,3*

1Department of Immunobiology, Medical Technology, and Science, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan.
2Department of Oncogenesis (C6), Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan.
3CREST, JST (Japan Science and Technology Corporation), Saitama 332-0012, Japan.

Abstract: The earliest biochemical events at the plasma membrane that lead to gene activation appear to depend not only on the local concentration of signaling molecules, but also on the mobility of these molecules at the site of signaling. To elucidate the process of signal transduction after receptor engagement in the immune system, it is important to analyze the mobility of signaling molecules in living lymphocytes. Current knowledge of the changes in intracellular localization and dynamic movements of signaling molecules during lymphocyte activation is limited. Here, we describe a method for known as fluorescence recovery after photobleaching, used to measure the diffusion mobility of a signaling molecule in a T cell line after T cell receptor stimulation. This method is a useful tool in studies of spatiotemporal regulation in immunoreceptor signaling.

*Corresponding author. Telephone, 81-6-6879-2599; e-mail: kosugi{at}sahs.med.osaka-u.ac.jp

Citation: N. Tanimura, M. Nagafuku, D. R. Liddicoat, T. Hamaoka, A. Kosugi, Analysis of the Mobility of Signaling Molecules in Lymphocytes Using Fluorescence Photobleaching Techniques. Sci. STKE 2003, pl10 (2003).

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