Human BDNF (Brain Derived Neurotrophic Factor) CLIA Kit-Laboratory Biological Research Reagents Manufacturer for Life Science - Elabscience

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Human BDNF (Brain Derived Neurotrophic Factor) CLIA Kit

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Price:   $682
Cat Number:  E-CL-H0010
Lead Time:   Order now, ship in 3 days

Reactivity:  Human

Detection Range:  78.13~5000 pg/mL

Sensitivity:  46.88 pg/mL

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Intended use

This CLIA kit applies to the in vitro quantitative determination of Human BDNF 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 Human BDNF. Standards or samples are added to the micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human BDNF 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 Human BDNF, 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 Human BDNF. You can calculate the concentration of Human BDNF in the samples by comparing the RLU value of the samples to the standard curve.

Assay type Sandwich-CLIA
Format 96T
Assay time 3.5h
Reactivity Human
Detection method Chemiluminescence
Detection range 78.13-5000 pg/mL
Sensitivity 46.88 pg/mL
Sample volume 100μL
Sample type serum, plasma and other biological fluids
Repeatability CV < 15%

Specificity

This kit recognizes Human BDNF in samples. No significant cross-reactivity or interference between Human BDNF 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
5000 59040
63472
61256 61233
2500 31634
37074
34354 34331
1250 18407
17931
18169 18146
625 9327
9457
9392 9369
312.5 5208
4458
4833 4810
156.25 2673
2349
2511 2488
78.13 1260
1418
1339 1316
0 22
24
23 --
CLIA-Elabscience

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human BDNF were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human BDNF 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) 275.32 499.95 2371.43 281.93 486.78 2574.49
Standard deviation 25.11 58.84 235.01 26.16 48.09 186.91
C V (%) 9.12 11.77 9.91 9.28 9.88 7.26

Recovery

The recovery of Human BDNF 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) 95-109 101
EDTA plasma (n=5) 94-112 102
Cell culture media (n=5) 98-111 103

Linearity

Samples were spiked with high concentrations of Human BDNF 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 (%) 100-114 97-114 92-106
Average (%) 108 105 97
1:4 Range (%) 92-108 98-111 88-100
Average (%) 100 105 95
1:8 Range (%) 99-116 91-106 93-108
Average (%) 107 98 99
1:16 Range (%) 87-103 89-101 88-102
Average (%) 94 95 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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