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A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
Authors:Darius J. R. Lane  Alfons Lawen
Affiliation:1.Molecular Pharmacology and Pathology Program, Department of Pathology & Bosch Institute, University of Sydney;2.Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University
Abstract:Vitamin C (ascorbate) plays numerous important roles in cellular metabolism, many of which have only come to light in recent years. For instance, within the brain, ascorbate acts in a neuroprotective and neuromodulatory manner that involves ascorbate cycling between neurons and vicinal astrocytes - a relationship that appears to be crucial for brain ascorbate homeostasis. Additionally, emerging evidence strongly suggests that ascorbate has a greatly expanded role in regulating cellular and systemic iron metabolism than is classically recognized. The increasing recognition of the integral role of ascorbate in normal and deregulated cellular and organismal physiology demands a range of medium-throughput and high-sensitivity analytic techniques that can be executed without the need for highly expensive specialist equipment. Here we provide explicit instructions for a medium-throughput, specific and relatively inexpensive microplate assay for the determination of both intra- and extracellular ascorbate in cell culture.
Keywords:Biochemistry   Issue 86   Vitamin C   Ascorbate   Cell swelling   Glutamate   Microplate assay   Astrocytes
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