Universal ELISA Kits

4-HNE(4-Hydroxynonenal) ELISA Kit

Product No

FY-EU1007

Sample Type

Serum, Plasma, Tissue homogenates, Cell lysates, Cell culture supernates and Other biological fluids

Assay Time

1.5H

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Product Details
Product Name in English:
4-HNE(4-Hydroxynonenal) ELISA Kit
Species:
Universal
Alternative Name:
4-HNE, HNE, 4-hydroxy-2-nonenal, 4-Hydroxynonenal
Assay Type:
Competitive
Standard:
40 ng/mL
Sensitivity:
0.38 ng/mL
Detection Range:
0.63-40 ng/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma, Tissue homogenates, Cell lysates, Cell culture supernates and Other biological fluids
Research:
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 stop 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.
Technical Data
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Concentration OD1 OD2 Average OD Corrected
40 0.251 0.253 0.252
20 0.414 0.416 0.415
10 0.654 0.66 0.657
5 0.971 0.973 0.972
2.5 1.264 1.268 1.266
1.25 1.584 1.599 1.592
0.63 1.854 1.865 1.86
0 2.676 2.687 2.682
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level were tested on 3 different plates, 20 replicates in each plate, respectively.
Item Intra-assay Precision Intra-assay Precision
Sample 3 3
Replicate 9 18
CV(%) 5 8
Rate of recovery
The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
Sample Type Range(%) Average Recovery(%)
Serum (n=8) 87-97 90
EDTA plasma (n=8) 81-95 88
Cell culture media (n=8) 83-95 89
Linear
Samples were spiked with high concentrations of target proteins and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2 Range (%) 95-103 85-94 82-90
Average (%) 96 89 88
1:4 Range (%) 86-92 85-94 85-92
Average (%) 90 94 91
1:8 Range (%) 89-97 80-96 80-96
Average (%) 92 95 95
1:16 Range (%) 91-103 92-101 92-101
Average (%) 91 98 96
Assay Procedures
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