Literature DB >> 21289434

Demonstration of a visual cell-based assay for screening glucose transporter 4 translocation modulators in real time.

Maleppillil Vavachan Vijayakumar1, Amrendra Kumar Ajay, Manoj Kumar Bhat.   

Abstract

Insulin-stimulated translocation of glucose transporter 4 (GLUT4) to cell membrane leading to glucose uptake is the rate-limiting step in diabetes. It is also a defined target of antidiabetic drug research. Existing GLUT4 translocation assays are based on time-consuming immunoassays and are hampered by assay variability and low sensitivity. We describe a real-time, visual, cell-based qualitative GLUT4 translocation assay using CHO-HIRc-myc-GLUT4eGFP cells that stably express myc- and eGFP-tagged GLUT4 in addition to human insulin receptor (HIRc). GLUT4 translocation is visualized by live cell imaging based on GFP fluorescence by employing a cooled charge-coupled device camera attached to a fluorescent microscope. This video imaging method and further quantitative analysis of GLUT4 on the cell membrane provide rapid and foolproof visual evidence that this method is suitable for screening GLUT4 translocation modulators.

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Year:  2010        PMID： 21289434     DOI： 10.1007/s12038-010-0060-0

Source DB:  PubMed          Journal:  J Biosci        ISSN： 0250-5991            Impact factor:   1.826


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