Rat IFNα (InterferonAlpha) CLIA Kit-Laboratory Biological Research Reagents Manufacturer for Life Science - Elabscience

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Rat IFNα (InterferonAlpha) CLIA Kit

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Price:   $682
Cat Number:  E-CL-R0753
Lead Time:   Order now, ship in 3 days

Reactivity:  Rat

Detection Range:  15.63~1000 pg/mL

Sensitivity:  9.38 pg/mL

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Intended use

This CLIA kit applies to the in vitro quantitative determination of Rat IFNα concentrations in serum, plasma and other biological fluids.

Test principle

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

Assay type Sandwich-CLIA
Format 96T
Assay time 4.5h
Reactivity Rat
Detection method Chemiluminescence
Detection range 15.63-1000 pg/mL
Sensitivity 9.38 pg/mL
Sample volume 100μL
Sample type serum, plasma and other biological fluids
Repeatability CV < 15%

Specificity

This kit recognizes Rat IFNα in samples. No significant cross-reactivity or interference between Rat IFNα 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
1000 34225
35503
34864 34791
500 17443
17123
17283 17210
250 7237
6813
7025 6952
125 3565
3723
3644 3571
62.5 1791
1985
1888 1815
31.25 901
873
887 814
15.63 387
351
369 296
0 77
69
73 --
sandwich-Ab-CLIA-Elabscience

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat IFNα were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat IFNα were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 49.94 120.79 432.76 45.45 105.60 453.74
Standard deviation 2.61 5.62 18.44 2.46 5.13 23.82
C V (%) 5.23 4.65 4.26 5.41 4.86 5.25

Recovery

The recovery of Rat IFNα 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) 91-98 94
EDTA plasma (n=5) 99-106 103
Cell culture media (n=5) 86-94 90

Linearity

Samples were spiked with high concentrations of Rat IFNα 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)
1:2 Range (%) 97-110 94-106 95-103
Average (%) 105 101 99
1:4 Range (%) 87-98 87-100 89-99
Average (%) 93 92 92
1:8 Range (%) 90-100 92-104 94-107
Average (%) 97 96 100
1:16 Range (%) 96-108 99-112 92-101
Average (%) 104 105 95

Kit Components& Storage

An unopened kit can be stored at 4℃ 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℃, 6 months
Reference Standard 2 vials
Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 μL
Concentrated HRP Conjugate (100×) 1 vial, 120 μL -20℃(shading light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 4℃, 6 months
Biotinylated Detection Ab Diluent 1 vial, 14 mL
HRP Conjugate Diluent 1 vial, 14 mL
Concentrated Wash Buffer (25×) 1 vial, 30 mL
Substrate Reagent A 1 vial, 5 mL 4℃ (shading light)
Substrate Reagent B 1 vial, 5 mL 4℃ (shading light)
Plate Sealer 5 pieces
Product Description 1 copy
Certificate of Analysis 1 copy

Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).

Other supplies required

  • Chemiluminescence immunoassay analyzer
  • High-precision transfer pipette, EP tubes and disposable pipette tips
  • Incubator capable of maintaining 37℃
  • Deionized or distilled water
  • Absorbent paper
  • Loading slot for Wash Buffer

Assay procedure

1.Add 100 μL of standard or sample to each well. Incubate for 90 min at 37℃.

2.Remove the liquid.

3.Add 100 μL Biotinylated Detection Ab. Incubate 1 hour at 37℃. Aspirate and wash 3 times.

4.Add 100 μL HRP Conjugate. Incubate 30 min at 37℃. Aspirate and wash 5 times.

5.Add 100 μL Substrate Mixture Solution. Incubate for 5 min at 37℃.

6.Fluorescence appeared. Measure the RLU value with the Chemiluminescence immunoassay analyzer. Calculation of the results.

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