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J Vis Exp. 2014 Apr 11;(86). doi: 10.3791/51322.

A rapid and specific microplate assay for the determination of intra- and extracellular ascorbate in cultured cells.

Author information

1
Molecular Pharmacology and Pathology Program, Department of Pathology & Bosch Institute, University of Sydney; darius.lane@sydney.edu.au.
2
Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University; alfons.lawen@monash.edu.

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.

PMID:
24747535
PMCID:
PMC4166944
DOI:
10.3791/51322
[Indexed for MEDLINE]
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