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Human HNE(4-Hydroxynonenal) ELISA Kit
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ELK0114
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48 T $320.00
96 T $458.00
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Overview

Product name: Human HNE(4-Hydroxynonenal) ELISA Kit
Reactivity: Human
Alternative Names: 4-HNE; HNE; 4-hydroxy-2-nonenal; 4-Hydroxynonenal
Assay Type: Sandwich
Sensitivity: 1.21 ng/mL
Standard: 200 ng/mL
Detection Range: 3.13-200 ng/mL
Sample type: Serum, plasma, tissue homogenates
Assay length: 3.5h
Research Area: Signal transduction;Metabolic pathway;Infection immunity;Hormone metabolism;Dermatology;
Test principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Human HNE. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human HNE. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Human HNE, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. 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 Human 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
200.00 2.045 1.977
100.00 1.562 1.494
50.00 1.285 1.217
25.00 0.915 0.847
12.50 0.634 0.566
6.25 0.389 0.321
3.13 0.258 0.190
0.00 0.068 0.000

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 HNE and the recovery rates were calculated by comparing the measured value to the expected amount of HNE in samples.
Matrix Recovery range Average
serum(n=5) 84-98% 91%
EDTA plasma(n=5) 79-97% 88%
Heparin plasma(n=5) 80-95% 87%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 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) 86-93% 91-105% 87-98% 89-94%
EDTA plasma(n=5) 95-103% 79-92% 97-105% 89-97%
Heparin plasma(n=5) 85-94% 97-106% 82-95% 93-105%