Rapid and sensitive determination of protein-nitrotyrosine by ELISA: Application to human plasma.

Abstract:

:3-Nitrotyrosine (3NT) is known as an important indicator of nitrosative stress and has been linked to various diseases. Our aim was to develop an indirect ELISA (enzyme-linked immunosorbent assay) method suitable for the detection of protein-bound 3NT in clinical plasma and serum samples. Nitrated protein standards and reduced protein standards were prepared. Limit of detection was determined for standards; recovery and reproducibility were determined for human plasma samples. The limit of detection for this method is 1.82±0.56 pmol/mg protein. Mean recovery of standards was 95%. 3NT concentration in plasma samples of obese and normal weight subjects was determined to be between 2 pmol/mg and 19 pmol/mg. No time-consuming sample preparation or expensive laboratory equipment is required, and applied antibodies are commercially available. Sensitivity, rapid analysis time, possibilities of high throughput applications and small sample volumes make this ELISA attractive for use in clinical laboratories.

journal_name

Free Radic Res

journal_title

Free radical research

authors

Weber D,Kneschke N,Grimm S,Bergheim I,Breusing N,Grune T

doi

10.3109/10715762.2011.652627

subject

Has Abstract

pub_date

2012-03-01 00:00:00

pages

276-85

issue

3

eissn

1071-5762

issn

1029-2470

journal_volume

46

pub_type

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