Isotope tracing enhancement of chemiluminescence assays for nitric oxide research
Isotope tracing enhancement of chemiluminescence assays for nitric oxide research
Chemiluminescence assays are used widely for the detection of nitric oxide (NO)-derived species in biological fluids and tissues. Here, we demonstrate that these assays can be interfaced with mass-sensitive detectors for parallel determination of isotopic abundance. Results obtained with tri-iodide and ascorbic acid-based reductive assays indicate that mass spectrometric detection enables NO isotope-tracing experiments to be carried out to a limit of detectability of a few picomoles, a sensitivity similar to that of standard gas phase chemiluminescence methods. The advantage afforded by mass spectrometric detection is demonstrated using the murine macrophage cell line J774, which is shown here to reduce 15NO3- to 15NO2- under anoxic conditions. The particular combination of an analytical and cellular system described here may hold promise for future characterization of the enzymatic pathways contributing to mammalian nitrate reductase activity, without background interference from 14NO2- derived from other sources.
181-189
Cornelius, Julia
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Tran, Tuan
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Turner, Nicole
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Piazza, Abigail
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Mills, Lauren
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Slack, Ryan
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Hauser, Sean
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Alexander, J. Steven
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Grisham, Matthew B.
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Feelisch, Martin
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Rodriguez, Juan
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February 2009
Cornelius, Julia
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Tran, Tuan
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Turner, Nicole
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Piazza, Abigail
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Mills, Lauren
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Slack, Ryan
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Hauser, Sean
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Alexander, J. Steven
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Grisham, Matthew B.
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Feelisch, Martin
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Rodriguez, Juan
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Cornelius, Julia, Tran, Tuan, Turner, Nicole, Piazza, Abigail, Mills, Lauren, Slack, Ryan, Hauser, Sean, Alexander, J. Steven, Grisham, Matthew B., Feelisch, Martin and Rodriguez, Juan
(2009)
Isotope tracing enhancement of chemiluminescence assays for nitric oxide research.
Biological Chemistry, 390 (2), .
(doi:10.1515/BC.2009.017).
(PMID:19040352)
Abstract
Chemiluminescence assays are used widely for the detection of nitric oxide (NO)-derived species in biological fluids and tissues. Here, we demonstrate that these assays can be interfaced with mass-sensitive detectors for parallel determination of isotopic abundance. Results obtained with tri-iodide and ascorbic acid-based reductive assays indicate that mass spectrometric detection enables NO isotope-tracing experiments to be carried out to a limit of detectability of a few picomoles, a sensitivity similar to that of standard gas phase chemiluminescence methods. The advantage afforded by mass spectrometric detection is demonstrated using the murine macrophage cell line J774, which is shown here to reduce 15NO3- to 15NO2- under anoxic conditions. The particular combination of an analytical and cellular system described here may hold promise for future characterization of the enzymatic pathways contributing to mammalian nitrate reductase activity, without background interference from 14NO2- derived from other sources.
Text
2008 Cornelius - Biol Chem - Just accepted.pdf_eprint.pdf
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Published date: February 2009
Organisations:
Clinical & Experimental Sciences
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Local EPrints ID: 337705
URI: http://eprints.soton.ac.uk/id/eprint/337705
ISSN: 1431-6730
PURE UUID: 899cdecf-aca1-4966-abae-484fa5d160fb
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Date deposited: 02 May 2012 13:16
Last modified: 15 Mar 2024 03:41
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Author:
Julia Cornelius
Author:
Tuan Tran
Author:
Nicole Turner
Author:
Abigail Piazza
Author:
Lauren Mills
Author:
Ryan Slack
Author:
Sean Hauser
Author:
J. Steven Alexander
Author:
Matthew B. Grisham
Author:
Juan Rodriguez
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