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Utilizing pHluorin-tagged Receptors to Monitor Subcellular Localization and Trafficking
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Utilizing pHluorin-tagged Receptors to Monitor Subcellular Localization and Trafficking

Article DOI: 10.3791/55466-v • 09:59 min • March 16th, 2017
March 16th, 2017 •
1, 2, 1
1Department of Chemistry, University of Kentucky, 2Department of Biomedical Sciences, Marshall University

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Summary
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Labeling the extracellular domain of a membrane protein with a pH sensitive fluorophore, superecliptic pHluorin (SEP), allows subcellular localization, expression, and trafficking to be determined. Imaging SEP-labeled proteins with total internal reflection fluorescence microscopy (TIRFM) enables the quantification of protein levels in the peripheral ER and plasma membrane.

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Fluorescence Imaging PHluorin-tagged Receptors Subcellular Localization Protein Trafficking Intracellular Distribution Membrane Protein Vesicle Delivery Protein Expression Real-time Imaging High Resolution Microscopy PH-sensitive Fluorophor Extracellular Solution TIRF Microscopy Epifluorescence Mode Exposure Time EM Gain Laser Beam
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