| Alternative Names: |
3-Nitro-L-Tyrosine; 3-Nitrotyrosine; Nitrotyrosine |
| Catalogue No. |
6687ELK |
| Size |
96T |
| Reactivity |
General |
| Range |
4.69-300 ng/mL |
| Sensitivity |
1.35 ng/mL |
| Assay Type |
Competitive Inhibition |
| 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 Nitrotyrosine(NT) protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Nitrotyrosine(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 Nitrotyrosine(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% |
|
|
For Research Use Only |
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