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ELK Biotechnology

ELK9448

General 10-HETE(10-Hydroxyeicosatetraenoic Acid) ELISA Kit

General 10-HETE(10-Hydroxyeicosatetraenoic Acid) ELISA Kit

Sensitivity

General 10-HETE(10-Hydroxyeicosatetraenoic Acid) ELISA Kit

Overview

Product name: 10-HETE(10-Hydroxyeicosatetraenoic Acid) ELISA Kit
Reactivity: General
Alternative Names: 10-HETE
Assay Type: Competitive Inhibition
Sensitivity: 3.95 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 10-HETE protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to 10-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 10-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 0.195 —
400.00 0.411 —
200.00 0.583 —
100.00 0.931 —
50.00 1.275 —
25.00 1.578 —
12.50 1.863 —
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 10-HETE and the recovery rates were calculated by comparing the measured value to the expected amount of 10-HETE in samples.
Matrix Recovery range Average
serum(n=5) 87-99% 93%
EDTA plasma(n=5) 90-105% 97%
Heparin plasma(n=5) 86-99% 92%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 10-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) 81-94% 93-107% 93-102% 91-105%
EDTA plasma(n=5) 85-97% 93-101% 97-103% 91-99%
Heparin plasma(n=5) 86-97% 92-101% 95-102% 98-106%
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