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Human 12-LO (Arachidonate 12-Lipoxygenase) CLIA Kit (HUES01277)

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SKU:
HUES01277
$681

Description

Human 12-LO (Arachidonate 12-Lipoxygenase) CLIA Kit

This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human 12-LO . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human 12-LO and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human 12-LO, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human 12-LO. The concentration of Human 12-LO in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection methodChemiluminescence
Detection range62.50-4000 pg/mL
Sensitivity37.50 pg/mL
Sample volume100uL
Sample typeTissue homogenates,cell lysates and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Human 12-LO in samples. No significant cross-reactivity or interference between Human 12-LO and analogues was observed.

Typical Data

As the RLU 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
(pg/mL)
RLU Average Corrected
400054424
63738
5908159046
200026225
29493
2785927824
100014621
12347
1348413449
5006433
6777
66056570
2503320
3166
32433208
1251629
1533
15811546
62.50742
768
755720
033
37
35--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human 12-LO were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human 12-LO were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)211.96500.641517.01213.47452.441571.15
Standard deviation26.1154.32134.2626.1550.31160.57
C V (%)12.3210.858.8512.2511.1210.22

Recovery

The recovery of Human 12-LO 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)88-10396
EDTA plasma (n=5)87-10093
Cell culture media (n=5)95-108102

Linearity

Samples were spiked with high concentrations of Human 12-LO 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 (%)88-10286-9695-112
Average (%)9491103
1:4Range (%)89-10486-9785-98
Average (%)969291
1:8Range (%)98-11592-10891-106
Average (%)10510098
1:16Range (%)97-11383-9594-109
Average (%)1059099

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 CLIA 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 Reagent A1 vial, 5 mL4'C (shading light)
Substrate Reagent B1 vial, 5 mL4'C (shading light)
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy
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Additional Information

Reactivity:
Human
ELISA Type:
CLIA
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