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3-NT(3-Nitrotyrosine) ELISA Kit
ELK7866
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48 T $265.00
96 T $376.00
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Overview

Product name: 3-NT(3-Nitrotyrosine) ELISA Kit
Reactivity: General
Alternative Names: 3-Nitro-L-Tyrosine; 3-Nitrotyrosine; Nitrotyrosine
Assay Type: Competitive Inhibition
Sensitivity: 1.35 ng/mL
Standard: 300 ng/mL
Detection Range: 4.69-300 ng/mL
Sample Type: serum, plasma and other biological fluids
Assay Length: 2h
Research Area: Metabolic pathway;Infection immunity;Rheumatology;
Test principle: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with 3-NT protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to 3-NT. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of 3-NT in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (ng/mL) OD Corrected OD
300.00 0.217
150.00 0.467
75.00 0.793
37.50 0.967
18.75 1.236
9.38 1.596
4.69 1.861
0.00 2.265

Precision

Intra-assay Precision (Precision within an assay)：CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays)：CV%<10%

Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.

Recovery

Matrices listed below were spiked with certain level of recombinant 3-NT and the recovery rates were calculated by comparing the measured value to the expected amount of 3-NT in samples.
Matrix Recovery range Average
serum(n=5) 92-107% 99%
EDTA plasma(n=5) 87-99% 93%
Heparin plasma(n=5) 78-92% 84%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 3-NT and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Matrix 1:2 1:4 1:8 1:16
serum(n=5) 86-93% 91-105% 88-104% 97-103%
EDTA plasma(n=5) 85-95% 79-96% 89-101% 85-97%
Heparin plasma(n=5) 87-96% 81-93% 86-98% 91-103%