The 5-HETE (5-Hydroxyeicosatetraenoic Acid) ELISA Kit is specifically designed for the accurate quantification of 5-HETE levels in biological samples such as serum, plasma, and tissue lysates. This kit offers high sensitivity and specificity, ensuring precise and reliable results for a variety of research applications.5-HETE is a biologically active lipid mediator derived from arachidonic acid metabolism that plays a key role in inflammation, immune response, and oxidative stress.
Abnormal levels of 5-HETE have been implicated in various inflammatory diseases and cancer, making it a valuable biomarker for studying these conditions and developing potential therapeutic interventions.With its user-friendly protocol and high-quality components, the 5-HETE ELISA Kit is an essential tool for researchers looking to investigate the role of 5-HETE in different pathophysiological processes and explore its potential as a target for novel treatment strategies.
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with 5-HETE. During the reaction, 5-HETE in the sample or standard competes with a fixed amount of 5-HETE on the solid phase supporter for sites on the Biotinylated Detection Ab specific to 5-HETE. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of 5-HETE in tested samples can be calculated by comparing the OD of the samples to the standard curve.
Linearity:
Serum (n=5)
EDTA plasma (n=5)
Cell culture media (n=5)
1:2
Range (%)
83-98
94-108
86-98
Average (%)
90
99
91
1:4
Range (%)
85-99
91-106
82-92
Average (%)
90
98
87
1:8
Range (%)
82-93
91-106
86-98
Average (%)
88
99
91
1:16
Range (%)
81-94
96-110
87-100
Average (%)
86
101
93
Recovery:
Sample Type
Range (%)
Average Recovery (%)
Serum (n=5)
95-107
101
EDTA plasma (n=5)
92-105
99
Cell culture media (n=5)
88-100
94
Precision:
Intra-assay Precision
Inter-assay Precision
Sample
1
2
3
1
2
3
n
20
20.0
20.0
20.0
20.0
20.0
Mean (ng/mL)
0.54
0.92
4.64
0.53
0.84
4.76
Standard deviation
0.04
0.05
0.16
0.03
0.04
0.18
C V (%)
7.41
5.43
3.45
5.66
4.76
3.87
Sample type &Sample volume:
Serum, plasma and other biological fluids; 50μL
Reproducibility:
Both intra-CV and inter-CV are < 10%.
Application:
This ELISA kit applies to the in vitro quantitative determination of 5-HETE concentrations in Serum, plasma and other biological fluids.
Specificity:
This kit recognizes 5-HETE in samples. No significant cross-reactivity or interference between 5-HETE and analogues was observed.