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ELK Biotechnology

ELK9581

General 4-HNE(4-Hydroxynonenal) ELISA Kit

General 4-HNE(4-Hydroxynonenal) ELISA Kit

Sensitivity

General 4-HNE(4-Hydroxynonenal) ELISA Kit

Overview

Product name: 4-HNE(4-Hydroxynonenal) ELISA Kit
Reactivity: General
Alternative Names: 4-HNE; HNE; 4-hydroxy-2-nonenal; 4-Hydroxynonenal
Assay Type: Competitive Inhibition
Sensitivity: 0.38 ng/mL
Standard: 40 ng/mL
Detection Range: 0.63-40 ng/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
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 4-HNE protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to 4-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 4-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
40.00 0.223 —
20.00 0.390 —
10.00 0.583 —
5.00 0.897 —
2.50 1.163 —
1.25 1.511 —
0.63 1.871 —
0.00 2.211 —

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) 95-107% 101%
EDTA plasma(n=5) 82-95% 88%
Heparin plasma(n=5) 90-105% 97%

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) 93-102% 80-93% 91-100% 89-97%
EDTA plasma(n=5) 86-102% 94-106% 93-101% 87-98%
Heparin plasma(n=5) 89-102% 98-102% 84-102% 83-95%
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