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5-HETE(5-Hydroxyeicosatetraenoic Acid) ELISA Kit (UNES00062)

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SKU:
UNES00062
€499

Description

5-HETE(5-Hydroxyeicosatetraenoic Acid) ELISA Kit

This ELISA kit uses Competitive-ELISA as the method. The microtiter 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 from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) are added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by adding Stop Solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of   5-HETE in the samples is then determined by comparing the OD of the samples to the standard curve.

Assay typeCompetitive-ELISA
Format96T
Assay time2.5h
ReactivityUniversal
Detection MethodColormetric
Detection Range0.16-10 ng/mL
Sensitivity0.09 ng/mL
Sample Volume50uL
Sample TypeSerum, 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.

Typical 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(ng/mL) O.D Average
100.321
0.375
0.348
50.455
0.465
0.46
2.50.67
0.644
0.657
1.250.957
0.963
0.96
0.631.343
1.333
1.338
0.311.733
1.723
1.728
0.162.011
2.015
2.013
02.428
2.442
2.435

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level 5-HETE were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level 5-HETE were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.540.924.640.530.844.76
Standard deviation0.040.050.160.030.040.18
C V (%)7.415.433.455.664.763.87

Recovery

The recovery of 5-HETE 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=5)95-107101
EDTA plasma (n=5)92-10599
Cell culture media (n=5)88-10094

Linearity

Samples were spiked with high concentrations of 5-HETE 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)
1:2Range (%)83-9894-10886-98
Average (%)909991
1:4Range (%)85-9991-10682-92
Average (%)909887
1:8Range (%)82-9391-10686-98
Average (%)889991
1:16Range (%)81-9496-11087-100
Average (%)8610193

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable)8 wells ×12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100×)1 vial, 120 uL
Concentrated HRP Conjugate (100×)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25×)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy
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Additional Information

Reactivity:
Universal
ELISA Type:
Competitive
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