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Robust Glucose and 1,5‐Anhydroglucitol Uptake Assays Using Novel Fluorescent Conjugates
Author(s) -
Li Shunan,
Khatwani Santoshkumar,
Lin Yuan,
Tchaga Grigority,
Yan Gordon
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.851.12
Subject(s) - conjugate , glucose uptake , glucose transporter , fluorescence , fluorescence microscope , biochemistry , chemistry , flow cytometry , glycolysis , citric acid cycle , carbohydrate metabolism , metabolism , biology , insulin , microbiology and biotechnology , endocrinology , mathematical analysis , physics , mathematics , quantum mechanics
Glucose is the primary carbon source for ATP production and biosynthesis in proliferating mammalian cells by either glycolytic or tricarboxylic acid (TCA) cycle pathways. These pathways are tightly regulated and their disruption leads to many metabolic diseases such as diabetes. Cancer cells exhibit increased glucose uptake to support their high rate of proliferation. The glucose uptake has been widely studied by a non‐radioactive assay using a fluorescently‐labeled glucose derivative, 2‐NBD‐Glucose. Despite its applicability, this conjugate exhibits photo bleaching and reduced fluorescence when used at low concentrations (i.e. <400 μM). BioVision has developed a novel bright green fluorescent 2‐deoxy‐glucose conjugate (GluTracker™) which, just like glucose and 2‐NBD‐Glucose, can be taken up by cells via glucose transporters. However, this glucose analog cannot be fully metabolized by cells, and thus accumulates inside cells. Fluorescence generated by this fluorescent glucose analog is proportional to the cellular glucose uptake and can be used to measure glucose uptake using fluorescence microscopy or flow cytometry. In addition, we have also developed a novel green fluorescent 1,5‐Anhydroglucitol conjugate (AGTracker TM ) to study the uptake of 1,5‐Anhdroglucitol (1,5‐AG). The blood plasma level of 1,5‐AG has been used in clinical studies to monitor short‐term glycemic control in diabetic patients. The AGTracker TM conjugate can be used to study 1,5‐AG uptake by fluorescence microscopy or flow cytometry. In conclusion, BioVision's GluTracker™ (for Glucose) and AGTracker TM (for 1,5‐AG) Uptake Assays provide valuable tools for drug discovery and identifying potential modulators in glucose‐dependent pathways. Support or Funding Information NA