| Alternative Names: |
4-HNE, HNE, 4-hydroxy-2-nonenal, 4-Hydroxynonenal |
| Catalogue No. |
3738ELK |
| Size |
96T |
| Reactivity |
Rat |
| Range |
15.63-1000 pg/mL |
| Sensitivity |
7.54 pg/mL |
| Assay Type |
Competitive Inhibition |
| Sample Type |
Serum, plasma, tissue homogenates |
| Assay Length |
2h |
| Research Area |
Signal transduction; Metabolic pathway; Infection immunity; Hormone metabolism; Dermatology; |
| Test principle |
This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with HNE protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to HNE. 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 HNE 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 |
| 1000.00 |
0.182 |
— |
| 500.00 |
0.275 |
— |
| 250.00 |
0.557 |
— |
| 125.00 |
0.897 |
— |
| 62.50 |
1.253 |
— |
| 31.25 |
1.587 |
— |
| 15.63 |
1.863 |
— |
| 0.00 |
2.475 |
— |
|
| 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 4-HNE and the recovery rates were calculated by comparing the measured value to the expected amount of 4-HNE in samples.
| Matrix |
Recovery range |
Average |
| serum(n=5) |
82-95% |
88% |
| EDTA plasma(n=5) |
95-107% |
101% |
| Heparin plasma(n=5) |
87-99% |
86% |
|
| Linearity |
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 4-HNE 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) |
82-93% |
95-104% |
87-96% |
85-97% |
| EDTA plasma(n=5) |
86-98% |
91-103% |
85-97% |
87-96% |
| Heparin plasma(n=5) |
83-94% |
83-96% |
92-102% |
82-93% |
|
| Note |
For Research Use Only |
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