12-HETE(12-Hydroxyeicosatetraenoic Acid) ELISA Kit (UNES00058)
12-HETE(12-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 12-HETE. During the reaction, 12-HETE in the sample or standard competes with a fixed amount of 12-HETE on the solid phase supporter for sites on the Biotinylated Detection Ab specific to 12-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 12-HETE in the samples is then determined by comparing the OD of the samples to the standard curve.
|Detection Range||0.78-50 ng/mL|
|Sample Type||Serum, plasma and other biological fluids|
This kit recognizes 12-HETE in samples. No significant cross-reactivity or interference between 12-HETE and analogues was observed.
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.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level 12-HETE were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level 12-HETE were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||5.65||5.26||4.15||4.93||4.52||3.34|
The recovery of 12-HETE spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||86-99||92|
|Cell culture media (n=5)||96-108||102|
Samples were spiked with high concentrations of 12-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)|
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.
|Micro ELISA Plate(Dismountable)||8 wells Ã12 strips||-20'C, 6 months|
|Reference Standard||2 vials|
|Concentrated Biotinylated Detection Ab (100Ã)||1 vial, 120 uL|
|Concentrated HRP Conjugate (100Ã)||1 vial, 120 uL||-20'C(shading light), 6 months|
|Reference Standard & Sample Diluent||1 vial, 20 mL||4'C, 6 months|
|Biotinylated Detection Ab Diluent||1 vial, 14 mL|
|HRP Conjugate Diluent||1 vial, 14 mL|
|Concentrated Wash Buffer (25Ã)||1 vial, 30 mL|
|Substrate Reagent||1 vial, 10 mL||4'C(shading light)|
|Stop Solution||1 vial, 10 mL||4'C|
|Plate Sealer||5 pieces|
|Product Description||1 copy|
|Certificate of Analysis||1 copy|