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

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

      Size:
      • 96T
      • 24T
      Qty:
      - +
      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

      Specificity

      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.

      Concentration
      (pg/mL)
      O.D Average Corrected
      500 2.427
      2.445
      2.436 2.376
      250 1.562
      1.584
      1.573 1.513
      125 0.993
      0.979
      0.986 0.926
      62.5 0.477
      0.515
      0.496 0.436
      31.25 0.245
      0.237
      0.241 0.181
      15.63 0.166
      0.16
      0.163 0.103
      7.81 0.105
      0.121
      0.113 0.053
      0 0.059
      0.061
      0.06 --

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

      Linearity

      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

      Citations

      1. Oxidative Burst Of Circulating Neutrophils Following Traumatic Brain Injury In Human.
        IF: 2.806
        Journal: Plos One(2013)
        DOI: 10.1371/journal.pone.0068963
        PMID: 23894384
        Products: E-EL-H0102 E-EL-H0109 
        Sample type: Plasma
        Reactivity: Human
      2. Decreased Monocyte Subsets And TLR4-Mediated Functions In Patients With Acute Severe Fever With Thrombocytopenia Syndrome (SFTS).
        IF: 2.532
        Journal: International Journal of Infectious Diseases(2015)
        DOI: 10.1016/j.ijid.2015.12.009
        PMID: 26701820
        Products: E-EL-H0109 E-EL-H0103 
        Sample type: Cell culture supernatant
        Reactivity: Human
      3. Inhibitory Effect Of miR-125b On Hepatitis C Virus Core Protein-Induced TLR2/MyD88 Signaling In THP-1 Macrophages.
        IF: 3.365
        Journal: World Journal of Gastroenterology(2016)
        DOI: 10.3748/wjg.v22.i17.4354
        PMID: 27158204
        Products: E-EL-H0109 E-EL-H0102 E-EL-H0103 
        Sample type: Cell culture supernatant
        Reactivity: Human
      4. Positive Feedback Cycle Of TNF-α Promotes Staphylococcal Enterotoxin B-Induced THP-1 Cell Apoptosis.
        IF: 4.3
        Journal: Frontier in Cellular and Infection Microbiology(2016)
        DOI: 10.3389/fcimb.2016.00109
        PMID: 27709104
        Products: E-EL-H0109 
        Sample type: Cell culture supernatant
        Reactivity: Human
      5. Multifunctional Dextran Sulfate-Coated Reconstituted High Density Lipoproteins Target Macrophages And Promote Beneficial Antiatherosclerotic Mechanisms.
        IF: 4.818
        Journal: Bioconjugate Chemistry(2016)
        DOI: 10.1021/acs.bioconjchem.6b00600
        PMID: 28004910
        Products: E-EL-H0109 E-EL-H0102 E-EL-H0103 
        Sample type: Cell culture supernatant
        Reactivity: Human
      6. Sublethal Effects Of Zinc Oxide Nanoparticles On Male Reproductive Cells.
        IF: 2.866
        Journal: Toxicology in Vitro(2016)
        DOI: 10.1016/j.tiv.2016.05.017
        PMID: 27247145
        Products: E-EL-H0109 
        Sample type: Cell culture supernatant
        Reactivity: Human
      7. The Role Of miR-19b In The Inhibition Of Endothelial Cell Apoptosis And Its Relationship With Coronary Artery Disease.
        IF: 4.259
        Journal: Scientific Reports(2015)
        DOI: 10.1038/srep15132
        PMID: 26459935
        Products: E-EL-H0109 E-EL-H0481 E-EL-H0661 
        Sample type: Serum
        Reactivity: Human
      8. In Vitro Antiproliferative, Pro-Apoptotic, Antimetastatic And Anti-Inflammatory Potential Of 2, 4-Diacteylphloroglucinol (DAPG) By Pseudomonas Aeruginosa Strain FP10.
        IF: 3.833
        Journal: Apoptosis(2015)
        DOI: 10.1007/s10495-015-1162-9
        PMID: 26283170
        Products: E-EL-H0109 E-EL-H0102 E-EL-H0149 
        Sample type: Cell culture supernatant
        Reactivity: Human
      9. 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.
        IF: 2.861
        Journal: Molecules(2015)
        DOI: 10.3390/molecules201219764
        PMID: 26633327
        Products: E-EL-H0102 E-EL-H0109 
        Sample type: Cell lysate
        Reactivity: Human
      10. C Reactive Protein And Enzymatically Modified LDL Cooperatively Promote Dendritic Cell-Mediated T Cell Activation.
        IF: 2.359
        Journal: Cardiovascular Pathology(2017)
        DOI: 10.1016/j.carpath.2017.03.009
        PMID: 28433861
        Products: E-EL-H0108 E-EL-H0109 E-EL-H0150 E-EL-H0102 
        Sample type: Cell culture supernatant
        Reactivity: Human
      11. Overcoming The Heat Endurance Of Tumor Cells By Interfering The Anaerobic Glycolysis Metabolism For Improved Photothermal Therapy.
        IF: 13.942
        Journal: Acs Nano(2017)
        DOI: 10.1021/acsnano.6b06658
        PMID: 28107631
        Products: E-EL-H0109 E-EL-H0102 E-EL-H0088 
        Sample type: Cell culture supernatant
        Reactivity: Human
      12. Ros-Dependent Activation Of Autophagy Through The PI3K/Akt/mTOR Pathway Is Induced By Hydroxysafflor Yellow A-Sonodynamic Therapy In THP-1 Macrophages.
        IF: 4.593
        Journal: Oxidative Medicine and Cellular Longevity(2017)
        DOI: 10.1155/2017/8519169
        PMID: 28191279
        Products: E-EL-H0109 E-EL-H0150 E-EL-H0149 
        Sample type: Cell culture supernatant
        Reactivity: Human
      13. The Pharmaceutical Device Prisma Skin Promotes In Vitro Angiogenesis Through Endothelial To Mesenchymal Transition During Skin Wound Healing.
        IF: 3.226
        Journal: International Journal of Molecular Sciences(2017)
        DOI: 10.3390/ijms18081614
        PMID: 28757565
        Products: E-EL-H0109 
        Sample type: Cell culture supernatant
        Reactivity: Human
      14. Zinc Oxide Nanoparticle-Induced Atherosclerotic Alterations In Vitro And In Vivo.
        IF: 4.3
        Journal: International Journal of Nanomedicine(2017)
        DOI: 10.2147/IJN.S134897
        PMID: 28652743
        Products: E-EL-H6008 E-EL-H0109 E-EL-H2172 E-EL-H1640 E-EL-R0580 E-EL-R0504 E-EL-R0488 E-EL-R0902 
        Sample type: Cell culture supernatant,Serum
        Reactivity: Human,Rat
      15. Inhibition Of miR27a Suppresses The Inflammatory Response Via The p38/MAPK Pathway In Intervertebral Disc Cells.
        IF: 1.261
        Journal: Experimental and Therapeutic Medicine(2017)
        DOI: 10.3892/etm.2017.5053
        PMID: 29067127
        Products: E-EL-H0149 E-EL-H0102 E-EL-H0109 
        Sample type: Cell culture supernatant
        Reactivity: Human
      16. High Anti-Human Cytomegalovirus Antibody Levels Are Associated With The Progression Of Essential Hypertension And Target Organ Damage In Han Chinese Population.
        IF: 2.806
        Journal: Plos One(2017)
        DOI: 10.1371/journal.pone.0181440
        PMID: 28837559
        Products: E-EL-H0043 E-EL-H0109 E-EL-H0102 
        Sample type: Serum
        Reactivity: Human
      17. Oleanolic acid protects against pathogenesis of atherosclerosis, possibly via FXR-mediated angiotensin (Ang)-(1-7) upregulation.
        IF: 2.759
        Journal: Biomedicine & Pharmacotherapy(2018)
        DOI: 10.1016/j.biopha.2017.11.151
        PMID: 29793333
        Products: E-EL-H5518 E-EL-H0755 E-EL-H0149 E-EL-H0109 E-EL-H0102 E-EL-RB0013 E-EL-RB0011 E-EL-RB0014 
        Sample type: Cell culture supernatant,Serum
        Reactivity: Human,Rabbit
      18. Role Of miR-589-3p In Human Lumbar Disc Degeneration And Its Potential Mechanism.
        IF: 1.261
        Journal: Experimental and Therapeutic Medicine(2018)
        DOI: 10.3892/etm.2017.5593
        PMID: 29434746
        Products: E-EL-H0109 E-EL-H0149 E-EL-H0102 
        Sample type: Cell culture supernatant
        Reactivity: Human
      19. The Shortening Telomere Length Of T Lymphocytes Maybe Associated With HyperFunction In Servere Aplastic Anemia.
        IF: 1.692
        Journal: Molecular Medicine Reports(2018)
        DOI: 10.3892/mmr.2017.8014
        PMID: 29115638
        Products: E-EL-H0108 E-EL-H0109 
        Reactivity: Human
      20. Tetrahydroxy Stilbene Glucoside Ameliorates H2O2-Induced Human Brain Microvascular Endothelial Cell Dysfunction In Vitro By Inhibiting Oxidative Stress And Inflammatory Responses.
        IF: 1.692
        Journal: Molecular Medicine Reports(2017)
        DOI: 10.3892/mmr.2017.7225
        PMID: 28849141
        Products: E-EL-H0109 E-EL-H0102 E-EL-H0149 
        Sample type: Cell culture supernatant
        Reactivity: Human
      21. Short-Term Postoperative Cognitive Dysfunction And Inflammatory Response In Patients Undergoing Cytoreductive Surgery And Hyperthermic Intraperitoneal Chemotherapy: A Pilot Study.
        IF: 3.232
        Journal: Mediators Of Inflammation(2017)
        DOI: 10.1155/2017/3605350
        PMID: 29249869
        Products: E-EL-H0149 E-EL-H0109 E-EL-H1554 E-EL-H2183 E-EL-H1297 
        Sample type: Serum
        Reactivity: Human
      22. Exploiting A New Strategy To Induce Immunogenic Cell Death To Improve Dendritic Cell-Based Vaccines For Lymphoma Immunotherapy.
        IF: 7.719
        Journal: OncoImmunology(2017)
        DOI: 10.1080/2162402X.2017.1356964
        PMID: 29147614
        Products: E-EL-H0109 
        Sample type: Cell culture supernatant
        Reactivity: Human
      23. Adiponectin may be used as a marker in prediction of diabetic retinopathy patients.
        Journal: Iraq Medical Journal(2018)
        Products: E-EL-H0109 
        Sample type: Serum
        Reactivity: Human
      24. Leukocyte Depletion Does Not Affect The In Vitro Healing Ability Of Platelet Rich Plasma.
        Journal: Experimental and Therapeutic Medicine(2018)
        DOI: 10.3892/etm.2018.5887
        PMID: 29556269
        Products: E-EL-H0108 E-EL-H1577 E-EL-H1575 E-EL-H1670 E-EL-0162 E-EL-H0109 E-EL-H0092 
        Reactivity: Human
      25. Biochemical investigation of gender-specific association between insulin resistance and inflammatory biomarkers in types 2 diabetic patients.
        IF: 3.457
        Journal: Biomedicine & Pharmacotherapy(2018)
        DOI: 10.1016/j.biopha.2018.06.044
        PMID: 29966972
        Products: E-EL-H0102 E-EL-H0109 E-EL-H0043 E-EL-H5439 
        Sample type: Serum
        Reactivity: Human
      26. High-Mobility Group Protein Box 1 is Upregulated in Children with Henoch-Schonlein Purpura.
        IF: 0.533
        Journal: Pediatric Allergy(2018)
        DOI: 10.1089/ped.2018.0872
        Products: E-EL-H1554 E-EL-H0102 E-EL-H6008 E-EL-H0109 
        Sample type: Serum
        Reactivity: Human
      27. Propofol Promotes Ankle Fracture Healing in Children by Inhibiting Inflammatory Response.
        IF: /
        Journal: Medical Science Monitor: International Medical Journal of Experimental And Clinical Research(2018)
        DOI: 10.12659/MSM.908592
        PMID: 29940605
        Products: E-EL-H0109 E-EL-H0149 E-EL-H0102 
        Sample type: Serum
        Reactivity: Human
      28. Serum Tnfα Levels Are Inversely Associated With Insulin Sensitivity in Diabetic Retinopathy Patients.
        Journal: Kerbala Jorunal of Medicine(2017)
        Products: E-EL-H0109 
        Sample type: Serum
        Reactivity: Human
      29. Potential Benefit of Melatonin as an Adjuvant Therapy in a Sample of Iraqi Patients with Crohn’s Disease: A New Study.
        Journal: ALMUSTANSIRIYAH Journal of PHARMACEUTICAL SCIENCES (AJPS)(2018)
        Products: E-EL-H0109 
        Sample type: Serum
        Reactivity: Human
      30. Tumor Necrosis Factor Alpha Synergetic with Insulin Resistance Potentially Contributes to the Development of Polycystic Ovary Syndrome.
        Journal: Medical Journal of Babylon(2017)
        Products: E-EL-H0109 
        Sample type: Serum
        Reactivity: Human
      31. Associations between knee structural measures, circulating inflammatory factors and MMP13 in patients with knee osteoarthritis.
        IF: 5.454
        Journal: Osteoarthritis and cartilage(2018)
        DOI: 10.1016/j.joca.2018.05.003
        PMID: 29753949
        Products: E-EL-H0109 E-EL-H0102 E-EL-H6008 E-EL-H5812 E-EL-H0253 
        Sample type: Serum
        Reactivity: Human
      32. Evaluation of the Anti-Inflammatory Effect of Telmisartan As an Adjuvant Therapy to NSAID in The Management of Knee Osteoarthritis; A Clinical Prospective Study.
        IF: /
        Journal: Iraqi JMS(2018)
        DOI: 10.22578/IJMS.16.1.14
        Products: E-EL-H0149 E-EL-H0109 E-EL-H5134 
        Sample type: Serum
        Reactivity: Human
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