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ELK Biotechnology

ELK8156

General 12-HETE(12-Hydroxyeicosatetraenoic Acid) ELISA Kit

General 12-HETE(12-Hydroxyeicosatetraenoic Acid) ELISA Kit

Size
Sensitivity
Detection range

General 12-HETE(12-Hydroxyeicosatetraenoic Acid) ELISA Kit

Overview

Product name: 12-HETE(12-Hydroxyeicosatetraenoic Acid) ELISA Kit
Reactivity: General
Alternative Names: 12HETE
Assay Type: Competitive Inhibition
Sensitivity: 5.32 ng/mL
Standard: 1000 ng/mL
Range: 15.63-1000 ng/mL
Sample Type: serum, plasma 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 12-HETE protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to 12-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 12-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
1000.00 0.175 —
500.00 0.352 —
250.00 0.655 —
125.00 0.833 —
62.50 1.197 —
31.25 1.596 —
15.63 1.789 —
0.00 2.115 —

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 12-HETE and the recovery rates were calculated by comparing the measured value to the expected amount of 12-HETE in samples.
Matrix Recovery range Average
serum(n=5) 87-97% 90%
EDTA plasma(n=5) 88-102% 95%
Heparin plasma(n=5) 80-93% 88%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 12-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) 88-97% 91-98% 91-103% 89-97%
EDTA plasma(n=5) 85-94% 97-106% 92-101% 82-90%
Heparin plasma(n=5) 95-102% 83-96% 88-95% 96-102%
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