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A chemiluminescense-based assay for S-nitrosoalbumin and other plasma S-nitrosothiols

A chemiluminescense-based assay for S-nitrosoalbumin and other plasma S-nitrosothiols
A chemiluminescense-based assay for S-nitrosoalbumin and other plasma S-nitrosothiols
The lack of a simple assay for the quantification of S-nitrosothiols in complex biological matrices has hampered our understanding of their contribution to normal physiology and pathophysiological states. In this paper we describe an assay based upon the release of nitric oxide by reaction with a mixture consisting of Cu(I), iodine and iodide with subsequent quantification by chemiluminescense. With this method we can detect levels of S-nitrosothiols down to 5 nM in plasma. Following alkylation of free thiols with N-ethylmaleimide, and removal of nitrite with acidified sulfanilamide, we were able to measure known amounts of S-nitrosoalbumin added to plasma or whole blood, with an inter-assay variation for plasma S-nitrosothiols of approximately 4%. Further studies showed that the mean concentration of circulating S-nitrosothiols in venous plasma of healthy human volunteers was 28+/-7 nM.
S-nitrosothiols, S-nitrosoalbumin, nitric oxide, chemiluminescence
1071-5762
1-9
Marley, Richard
fd35dd15-d96d-42d2-a1f6-67ed55636deb
Feelisch, Martin
8c1b9965-8614-4e85-b2c6-458a2e17eafd
Holt, Stephen
ced68993-d00d-4778-b5cb-409f0c354d2e
Moore, Kevin
0f0c315d-93bd-4231-8ad1-aa11ac9dbf10
Marley, Richard
fd35dd15-d96d-42d2-a1f6-67ed55636deb
Feelisch, Martin
8c1b9965-8614-4e85-b2c6-458a2e17eafd
Holt, Stephen
ced68993-d00d-4778-b5cb-409f0c354d2e
Moore, Kevin
0f0c315d-93bd-4231-8ad1-aa11ac9dbf10

Marley, Richard, Feelisch, Martin, Holt, Stephen and Moore, Kevin (2000) A chemiluminescense-based assay for S-nitrosoalbumin and other plasma S-nitrosothiols. Free Radical Research, 32 (1), 1-9. (doi:10.1080/10715760000300011). (PMID:10625212)

Record type: Article

Abstract

The lack of a simple assay for the quantification of S-nitrosothiols in complex biological matrices has hampered our understanding of their contribution to normal physiology and pathophysiological states. In this paper we describe an assay based upon the release of nitric oxide by reaction with a mixture consisting of Cu(I), iodine and iodide with subsequent quantification by chemiluminescense. With this method we can detect levels of S-nitrosothiols down to 5 nM in plasma. Following alkylation of free thiols with N-ethylmaleimide, and removal of nitrite with acidified sulfanilamide, we were able to measure known amounts of S-nitrosoalbumin added to plasma or whole blood, with an inter-assay variation for plasma S-nitrosothiols of approximately 4%. Further studies showed that the mean concentration of circulating S-nitrosothiols in venous plasma of healthy human volunteers was 28+/-7 nM.

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More information

Published date: January 2000
Keywords: S-nitrosothiols, S-nitrosoalbumin, nitric oxide, chemiluminescence
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 337878
URI: https://eprints.soton.ac.uk/id/eprint/337878
ISSN: 1071-5762
PURE UUID: ae6c4fd0-43b1-4514-b46a-5283e116d03e
ORCID for Martin Feelisch: ORCID iD orcid.org/0000-0003-2320-1158

Catalogue record

Date deposited: 29 Jun 2012 10:56
Last modified: 06 Jun 2018 12:29

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