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Rat 5-LO (Arachidonate 5-Lipoxygenase) CLIA Kit (RTES00563)

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SKU:
RTES00563
$738

Description

Rat 5-LO (Arachidonate 5-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 Rat 5-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 Rat 5-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 Rat 5-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 Rat 5-LO. The concentration of Rat 5-LO in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityRat
Detection methodChemiluminescence
Detection range31.25-2000 pg/mL
Sensitivity18.75 pg/mL
Sample volume100uL
Sample typeTissue homogenates,cell lysates and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Rat 5-LO in samples. No significant cross-reactivity or interference between Rat 5-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
200054978
56464
5572155693
100023097
24851
2397423946
50011134
10930
1103211004
2504846
5742
52945266
1252609
2607
26082580
62.51412
1210
13111283
31.25662
686
674646
027
29
28--

Precision

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

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

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)101.62240.31769.41103.52245.77798.37
Standard deviation8.8019.3254.328.9823.8490.77
C V (%)8.668.047.068.679.7011.37

Recovery

The recovery of Rat 5-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)90-10496
EDTA plasma (n=5)95-112103
Cell culture media (n=5)87-10093

Linearity

Samples were spiked with high concentrations of Rat 5-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 (%)84-10098-11586-101
Average (%)9110693
1:4Range (%)92-10690-10490-101
Average (%)989596
1:8Range (%)100-112102-11787-100
Average (%)10610992
1:16Range (%)102-11898-11397-111
Average (%)110104103

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:
Rat
ELISA Type:
CLIA
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