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An assay for the rate of removal of extracellular hydrogen peroxide by cells
Journal article   Open access   Peer reviewed

An assay for the rate of removal of extracellular hydrogen peroxide by cells

Brett A Wagner, Jordan R Witmer, Thomas J van't Erve and Garry R Buettner
Redox biology, Vol.1(1), pp.210-217
2013
DOI: 10.1016/j.redox.2013.01.011
PMCID: PMC3736862
PMID: 23936757
url
https://doi.org/10.1016/j.redox.2013.01.011View
Published (Version of record) Open Access

Abstract

Cells have a wide range of capacities to remove extracellular hydrogen peroxide. At higher concentrations of extracellular H 2 O 2 (micromolar) the rate of removal can be approximated by a rate equation that is first-order in the concentration of H 2 O 2 and cell density. Here we present a method to determine the observed rate constant for the removal of extracellular H 2 O 2 on a per cell basis. In the cells examined, when exposed to 20 μM H 2 O 2 , these rate constants ( k cell ) range from 0.46×10 −12  s −1  cell −1  L for Mia-PaCa-2 cells (human pancreatic carcinoma) to 10.4×10 −12  s −1  cell −1  L for U937 cells (human histiocytic lymphoma). For the relatively small red blood cell k cell =2.9×10 −12  s −1  cell −1  L. These rate constants, k cell , can be used to compare the capacity of cells to remove higher levels of extracellular H 2 O 2 , as often presented in cell culture experiments. They also provide a means to estimate the rate of removal of extracellular H 2 O 2 , rate=− k cell [H 2 O 2 ] (cells L −1 ), and the half-life of a bolus of H 2 O 2 . This information is essential to optimize experimental design and interpret data from experiments that expose cells to extracellular H 2 O 2 .
Erythrocyte Hydrogen peroxide Quantitative redox biology RBC, Red blood cells Method Kinetics QRB, Quantitative Redox Biology

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