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Human TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit– MSE Supplies LLC

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Human TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit

SKU: E-EL-H0109

  • £32800
  • Save £5900



Human TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit

Detection Range 7.81-500 pg/mL
Sensitivity 4.69 pg/mL

 

Product Details

Properties

Assay type Sandwich-ELISA
Format 96T
Assay time 3.5h
Detection range 7.81-500 pg/mL
Sensitivity 4.69 pg/mL
Sample type &Sample volume serum, plasma and other biological fluids; 100μL
Specificity This kit recognizes Human TNF-α in samples. No significant cross-reactivity or interference between Human TNF-α and analogues was observed.
Reproducibility Both intra-CV and inter-CV are < 10%.
Application This ELISA kit applies to the in vitro quantitative determination of Human TNF-α concentrations in serum, plasma and other biological fluids.


Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human TNF-α. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human TNF-α 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 TNF-α, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human TNF-α. You can calculate the concentration of Human TNF-α in the samples by comparing the OD of the samples to the standard curve.

 

Kit Components and Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

    Item Specifications Storage
    Micro ELISA Plate(Dismountable) 96T: 8 wells ×12 strips
    48T: 8 wells ×6 strips
    -20℃, 6 months
    Reference Standard 96T: 2 vials
    48T: 1 vial
    Concentrated Biotinylated Detection Ab (100×) 96T: 1 vial, 120 μL
    48T: 1 vial, 60 μL
    Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL
    48T: 1 vial, 60 μL
    -20℃(Protect from light), 6 months
    Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 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 1 vial, 10 mL 2-8℃(Protect from light)
    Stop Solution 1 vial, 10 mL 2-8°C
    Plate Sealer 5 pieces
    Manual 1 copy
    Certificate of Analysis 1 copy

     

     

    Technical Data:

    Precision

    Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human TNF-α were tested 20 times on one plate, respectively.
    Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human TNF-α 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) 24.10 79.50 183.00 22.70 86.50 177.40
    Standard deviation 1.50 3.80 9.70 1.20 4.00 7.60
    CV (%) 6.22 4.78 5.30 5.29 4.62 4.28

     

     Recovery

    The recovery of Human TNF-α 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=8) 86-100 92
    EDTA plasma (n=8) 94-106 99
    Cell culture media (n=8) 93-107 99


    Target Information

    Database Links  SwissProt: P01375
    Synonyms DIF, TNF-alpha, TNFA, TNFSF2
    Research Area Cancer, Cardiovascular, Metabolism, Immunology, Signal transduction

     

     Assay Procedures

    elisa assay procedure 1

    1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C

    elisa assay procedure 2

    2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C

    elisa assay procedure 3

    3. Aspirate and wash the plate for 3 times

    elisa assay procedure 4

    4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times

    elisa assay procedure 5

    5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

    elisa assay procedure 6

    6. Add 50μL Stop Solution

    elisa assay procedure 7

    7. Read the plate at 450nm immediately. Calculation of the results