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


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).

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Description/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.

Item Type: Article
Digital Object Identifier (DOI): doi:10.1080/10715760000300011
ISSNs: 1071-5762 (print)
1029-2470 (electronic)
Keywords: S-nitrosothiols, S-nitrosoalbumin, nitric oxide, chemiluminescence
Subjects: Q Science > QP Physiology
Q Science > QR Microbiology > QR180 Immunology
Divisions : Faculty of Medicine > Clinical and Experimental Sciences
ePrint ID: 337878
Accepted Date and Publication Date:
Status
January 2000Published
Date Deposited: 29 Jun 2012 10:56
Last Modified: 31 Mar 2016 14:27
URI: http://eprints.soton.ac.uk/id/eprint/337878

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