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5-HETE(5-Hydroxyeicosatetraenoic Acid) ELISA Kit
ELK8155
Size： Price：
48 T $265.00
96 T $376.00
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Overview

Product name: 5-HETE(5-Hydroxyeicosatetraenoic Acid) ELISA Kit
Reactivity: General
Alternative Names: 5-HETE
Assay Type: Competitive Inhibition
Sensitivity: 3.83 ng/mL
Standard: 800 ng/mL
Detection Range: 12.5-800 ng/mL
Sample Type: Serum, plasma and other biological fluids
Assay Length: 2h
Research Area: Enzyme & Kinase;
Test principle: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with 5-HETE. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to 5-HETE. 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 5-HETE 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
800.00 0.191
400.00 0.367
200.00 0.577
100.00 0.861
50.00 1.119
25.00 1.578
12.50 1.775
0.00 2.123

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 5-HETE and the recovery rates were calculated by comparing the measured value to the expected amount of 5-HETE in samples.
Matrix Recovery range Average
serum(n=5) 78-91% 84%
EDTA plasma(n=5) 88-101% 94%
Heparin plasma(n=5) 82-95% 97%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 5-HETE 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) 97-106% 86-92% 81-93% 89-97%
EDTA plasma(n=5) 90-103% 97-106% 82-90% 92-101%
Heparin plasma(n=5) 82-95% 83-95% 88-94% 96-101%