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Rab GTPases pp 173-179 | Cite as

Selective Visualization of GLUT4 Storage Vesicles and Associated Rab Proteins Using IRAP-pHluorin

  • Yu Chen
  • Jennifer Lippincott-SchwartzEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1298)

Abstract

Fluorescence microscopy and fluorescent protein (FP)-tagged GLUT4 molecule have been great tools to characterize GLUT4 localization and dynamics inside the cell. However, it was difficult to distinguish GLUT4 storage vesicles (GSVs) from other intracellular compartments containing GLUT4 in live cells. Here, we describe the use of IRAP-pHluorin and total internal reflection fluorescence (TIRF) microscopy to selectively visualize GSVs and Rab proteins that associate with GSVs. This assay is also valuable to further defining GSV identity by unraveling other GSV-associated proteins.

Key words

GLUT4 IRAP Rab10 AS160 TIRF Adipocytes 

Notes

Acknowledgement

We thank all members of J.L.-S.’s group for helpful discussions.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Cell Biology and Metabolism Program, NICHD – Eunice Kennedy Shriver National Institute of Child Health and Human DevelopmentNIH – National Institutes of HealthBethesdaUSA

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