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FACS-based Glucose Uptake Assay of Mouse Embryonic Fibroblasts and Breast Cancer Cells Using 2-NBDG Probe

Overview
Journal Bio Protoc
Specialty Biology
Date 2021 Jul 21
PMID 34286029
Citations 3
Authors
Shengli Dong
Suresh K Alahari
Affiliations
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Abstract

This is a flow cytometry-based protocol to measure glucose uptake of mouse embryonic fibroblasts (MEFs) and breast cancer cells . The method is a slightly modified and updated version as previously described ( Dong , 2017 ). Briefly, the target cells are incubated with the fluorescently tagged 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG) for 2 h or 30 min, and the efficiency of glucose uptake is examined using a flow cytometer. This method can be adapted to measure a variety of adipocytes, immune cells, MEFs and cancer cells.

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References
1.
Dong S, Maziveyi M, Alahari S . Primary Tumor and MEF Cell Isolation to Study Lung Metastasis. J Vis Exp. 2015; (99):e52609. PMC: 4542731. DOI: 10.3791/52609. View
2.
Nitin N, Carlson A, Muldoon T, El-Naggar A, Gillenwater A, Richards-Kortum R . Molecular imaging of glucose uptake in oral neoplasia following topical application of fluorescently labeled deoxy-glucose. Int J Cancer. 2009; 124(11):2634-42. PMC: 2700039. DOI: 10.1002/ijc.24222. View
3.
ONeil R, Wu L, Mullani N . Uptake of a fluorescent deoxyglucose analog (2-NBDG) in tumor cells. Mol Imaging Biol. 2005; 7(6):388-92. DOI: 10.1007/s11307-005-0011-6. View
4.
Vander Heiden M, Cantley L, Thompson C . Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science. 2009; 324(5930):1029-33. PMC: 2849637. DOI: 10.1126/science.1160809. View
5.
Zou C, Wang Y, Shen Z . 2-NBDG as a fluorescent indicator for direct glucose uptake measurement. J Biochem Biophys Methods. 2005; 64(3):207-15. DOI: 10.1016/j.jbbm.2005.08.001. View