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

    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience

      Catalog number:E-EL-H0109

      Synonyms:DIF, TNF-alpha, TNFA, TNFSF2

      • 96T
      • 24T
      - +
      Price: $495

      Reactivity: Human

      Detection Range: 7.81~500 pg/mL

      Sensitivity: 4.69 pg/mL

      Lead Time: 7~10 daysWelcome to order from local distributors.

      Add to cart Compare Bulk request

      Intended use

      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.

      Assay type Sandwich
      Format 96T
      Assay time 3.5h
      Reactivity Human
      Detection Method Colormetric
      Detection Range 7.81—500 pg/mL
      Sensitivity 4.69 pg/mL
      Sample Volume Required Per Well 100μL
      Sample Type Serum, plasma and other biological fluids


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

      Typical data

      As the OD 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.

      O.D Average Corrected
      500 2.427
      2.436 2.376
      250 1.562
      1.573 1.513
      125 0.993
      0.986 0.926
      62.5 0.477
      0.496 0.436
      31.25 0.245
      0.241 0.181
      15.63 0.166
      0.163 0.103
      7.81 0.105
      0.113 0.053
      0 0.059
      0.06 --


      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
      C V (%) 6.22 4.78 5.30 5.29 4.62 4.28


      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=5) 86-100 92
      EDTA plasma (n=5) 94-106 99
      Cell culture media (n=5) 93-107 99


      Samples were spiked with high concentrations of Human TNF-α 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 (%) 92-105 84-97 88-99
      Average (%) 99 91 93
      1:4 Range (%) 99-116 87-99 91-104
      Average (%) 107 93 98
      1:8 Range (%) 97-113 81-93 92-109
      Average (%) 104 88 99
      1:16 Range (%) 96-112 85-98 95-107
      Average (%) 103 90 100

      Kit components & Storage

      An unopened kit can be stored at 2-8°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 ELISA Plate(Dismountable) 8 wells ×12 strips -20°C, 6 months
      Reference Standard 2 vials
      Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 μL
      Concentrated HRP Conjugate (100×) 1 vial, 120 μL -20°C(shading 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°C(shading light)
      Stop Solution 1 vial, 10 mL 2-8°C
      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

      • Microplate reader with 450 nm wavelength filter
      • High-precision transfer pipette, EP tubes and disposable pipette tips
      • Incubator capable of maintaining 37°C
      • Deionized or distilled water
      • Absorbent paper
      • Loading slot for Wash Buffer

      Assay Procedure

      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


      1. Publication: Liao Y, Liu P, Guo F, et al. Oxidative Burst Of Circulating Neutrophils Following Traumatic Brain Injury In Human[J]. Plos One, 2013.
        Species: Human
        Sample Type: Plasma
      2. Publication: Cheng P, Hua W, Zhang W, et al. Decreased Monocyte Subsets And TLR4-Mediated Functions In Patients With Acute Severe Fever With Thrombocytopenia Syndrome (SFTS)[J]. International Journal of Infectious Diseases, 2015, 43: 37-42.
        Species: Human
        Sample Type: Culture supernatant
      3. Publication: Cheng P, Hua W, Zhang W J, et al. Inhibitory Effect Of miR-125b On Hepatitis C Virus Core Protein-Induced TLR2/MyD88 Signaling In THP-1 Macrophages[J]. World Journal of Gastroenterology, 2016, 22(17): 4354-4361.
        Species: Human
        Sample Type: Culture supernatant
      4. Publication: Zhang X P, Shang W L, Yuan J Z, et al. Positive Feedback Cycle Of TNF-α Promotes Staphylococcal Enterotoxin B-Induced THP-1 Cell Apoptosis[J]. Frontier in Cellular and Infection Microbiology, 2016.
        Species: Human
        Sample Type: Culture supernatant
      5. Publication: Zhao Y, Jiang C, He J, et al. Multifunctional Dextran Sulfate-Coated Reconstituted High Density Lipoproteins Target Macrophages And Promote Beneficial Antiatherosclerotic Mechanisms[J]. Bioconjugate Chemistry, 2016.
        Species: Human
        Sample Type: Culture supernatant
      6. Publication: Liu Q, Xu C, Ji G, et al. Sublethal Effects Of Zinc Oxide Nanoparticles On Male Reproductive Cells[J]. Toxicology in Vitro, 2016, 35: 131-138.
        Species: Human
        Sample Type: Culture supernatant
      7. Publication: Tang Y, Zhang Y C, Chen Y, et al. The Role Of miR-19b In The Inhibition Of Endothelial Cell Apoptosis And Its Relationship With Coronary Artery Disease[J]. Scientific Reports, 2015.
        Species: Human
        Sample Type: Serum
      8. Publication: Veena V K, Popavath R N, Kennedy K, et al. In Vitro Antiproliferative, Pro-Apoptotic, Antimetastatic And Anti-Inflammatory Potential Of 2, 4-Diacteylphloroglucinol (DAPG) By Pseudomonas Aeruginosa Strain FP10[J]. Apoptosis, 2015, 20(10): 1281-1295.
        Species: Human
        Sample Type: Culture supernatant
      9. Publication: Hao S Y, Ji J J, Zhao H T,et al. Mitochondrion-Targeted Peptide SS-31 Inhibited Oxidized Low-Density Lipoproteins-Induced Foam Cell Formation Through Both ROS Scavenging And Inhibition Of Cholesterol Influx In Raw264.7 Cells[J]. Molecules, 2015, 20(12): 21287-21297.
        Species: Human
        Sample Type: Cell lysis
      10. Publication: He W, Ren Y, Wang X, et al. C Reactive Protein And Enzymatically Modified LDL Cooperatively Promote Dendritic Cell-Mediated T Cell Activation[J]. Cardiovascular Pathology, 2017, 29: 1-6.
        Species: Human
        Sample Type: Culture supernatant
      11. Publication: Chen W H, Luo G F, Lei Q, et al. Overcoming The Heat Endurance Of Tumor Cells By Interfering The Anaerobic Glycolysis Metabolism For Improved Photothermal Therapy[J]. Acs Nano, 2017, 11(2): 1419-1431.
        Species: Human
        Sample Type: Culture supernatant
      12. Publication: Jiang Y, Kou J, Han X, et al. Ros-Dependent Activation Of Autophagy Through The PI3K/Akt/mTOR Pathway Is Induced By Hydroxysafflor Yellow A-Sonodynamic Therapy In THP-1 Macrophages[J]. Oxidative Medicine and Cellular Longevity, 2017.
        Species: Human
        Sample Type: Culture supernatant
      13. Publication: Belvedere R, Bizzarro V, Parente L, et al. The Pharmaceutical Device Prisma® Skin Promotes In Vitro Angiogenesis Through Endothelial To Mesenchymal Transition During Skin Wound Healing[J]. International Journal of Molecular Sciences, 2017.
        Species: Human
        Sample Type: Culture supernatant
      14. Publication: Yan Z, Wang W, Wu Y, et al. Zinc Oxide Nanoparticle-Induced Atherosclerotic Alterations In Vitro And In Vivo[J]. International Journal of Nanomedicine, 2017, 12: 4433-4442.
        Species: Human, Rat
        Sample Type: Culture supernatant and Serum
      15. Publication: Cao Z, Chen L. Inhibition Of miR‑27a Suppresses The Inflammatory Response Via The p38/MAPK Pathway In Intervertebral Disc Cells[J]. Experimental and Therapeutic Medicine, 2017, 14(5): 4572-4578.
        Species: Human
        Sample Type: Culture supernatant
      16. Publication: Li Z, Tang Y, Tang N, et al. High Anti-Human Cytomegalovirus Antibody Levels Are Associated With The Progression Of Essential Hypertension And Target Organ Damage In Han Chinese Population[J]. Plos One, 2017.
        Species: Human
        Sample Type: Serum
      17. Publication: Pan Y Y, Zhou F H, Song Z H, et al. Oleanolic acid protects against pathogenesis of atherosclerosis, possibly via FXR-mediated angiotensin (Ang)-(1-7) upregulation[J]. Biomedicine & Pharmacotherapy, 2018, 97: 1694-1700.
        Species: Human, Rabbit
        Sample Type: Serum and culture supernatant
      18. Publication: Lu A Q, Wang Z R, Wang S C. Role Of miR-589-3p In Human Lumbar Disc Degeneration And Its Potential Mechanism[J]. Experimental and Therapeutic Medicine, 2018, 15: 1616-1621.
        Species: Human
        Sample Type: Culture supernatant
      19. Publication: Wang C M, Zhang T, Wang Y H, et al. The Shortening Telomere Length Of T Lymphocytes Maybe Associated With Hyper‑Function In Servere Aplastic Anemia[J]. Molecular Medicine Reports, 2018, 17: 1015-1021.
        Species: Human
      20. Publication: Jiang Z, Wang W H, Guo C C. Tetrahydroxy Stilbene Glucoside Ameliorates H2O2-Induced Human Brain Microvascular Endothelial Cell Dysfunction In Vitro By Inhibiting Oxidative Stress And Inflammatory Responses[J]. Molecular Medicine Reports, 2017, 16(4): 5219-5224.
        Species: Human
        Sample Type: Culture supernatant
      21. Publication: Yu H, Dong R, Lu Y Y, et al. Short-Term Postoperative Cognitive Dysfunction And Inflammatory Response In Patients Undergoing Cytoreductive Surgery And Hyperthermic Intraperitoneal Chemotherapy: A Pilot Study[J]. Mediators Of Inflammation, 2017.
        Species: Human
        Sample Type: Serum
      22. Publication: Montico B, Lapenta C, Ravo M, et al. Exploiting A New Strategy To Induce Immunogenic Cell Death To Improve Dendritic Cell-Based Vaccines For Lymphoma Immunotherapy[J]. OncoImmunology, 2017.
        Species: Human
        Sample Type: Culture supernatant
      23. Publication: Hatef Z S, Nada S Z, Jasim A M. Adiponectin may be used as a marker in prediction of diabetic retinopathy patients[J]. Iraq Medical Journal, 2018, 2(1): 20-23.
      24. Publication: Giusti I, Di Francesco M D, D'Ascenzo S, et al. Leukocyte Depletion Does Not Affect The In Vitro Healing Ability Of Platelet Rich Plasma[J]. Experimental and Therapeutic Medicine, 2018, 15(4): 4029-4038.
        Species: Human
      25. Publication: Akash M S H, Rehman K, Liaqat A, et al. Biochemical investigation of gender-specific association between insulin resistance and inflammatory biomarkers in types 2 diabetic patients[J]. Biomedicine & Pharmacotherapy, 2018, 106: 285-291.
        Species: Human
        Sample Type: Serum
      26. Publication: Wang F, Li X, Huang L, et al. High-Mobility Group Protein Box 1 is Upregulated in Children with Henoch-Schonlein Purpura[J]. Pediatric Allergy, Immunology, and Pulmonology, 2018, 31(2): 66-72.
        Species: Human
        Sample Type: Serum
      27. Publication: Zhou P, Liu H, Wu Y, et al. Propofol Promotes Ankle Fracture Healing in Children by Inhibiting Inflammatory Response[J]. Medical Science Monitor: International Medical Journal of Experimental And Clinical Research, 2018, 24: 4379.
        Species: Human
        Sample Type: Serum
      28. Publication: Nada S Z, Hatef Z S, Alkutubi H M, et al. Serum Tnfα Levels Are Inversely Associated With Insulin Sensitivity in Diabetic Retinopathy Patients[J]. Kerbala Jorunal of Medicine, 2017, 10(3): 3857-3862.
        Species: Human
        Sample Type: Serum
      29. Publication: Saleh H H, Abdulridha M K, Najeb A A. Potential Benefit of Melatonin as an Adjuvant Therapy in a Sample of Iraqi Patients with Crohn’s Disease: A New Study[J]. ALMUSTANSIRIYAH Journal of PHARMACEUTICAL SCIENCES (AJPS), 2018, 17(2): 9-9.
        Species: Human
        Sample Type: Serum
      30. Publication: Al-Assadi N A M, Al-Qaysi S A. Tumor Necrosis Factor Alpha Synergetic with Insulin Resistance Potentially Contributes to the Development of Polycystic Ovary Syndrome[J]. Medical Journal of Babylon, 2017, 14(4): 732-737.
        Species: Human
        Sample Type: Serum
      31. Publication: Ruan G, Xu J, Wang K, et al. Associations between knee structural measures, circulating inflammatory factors and MMP13 in patients with knee osteoarthritis[J]. Osteoarthritis and cartilage, 2018.
        Species: Human
        Sample Type: Serum
      32. Publication: Jasim N A, Kamal Y M, Mohammed M M, et al. 14. Evaluation of the Anti-Inflammatory Effect of Telmisartan As an Adjuvant Therapy to NSAID in The Management of Knee Osteoarthritis; A Clinical Prospective Study[J]. Iraqi JMS 2018; Vol. 16(1): 100-110.
        Species: Human
        Sample Type: Serum
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