• Product name

    Human TNF alpha ELISA Kit
    See all TNF alpha kits
  • Detection method

  • Sample type

    Cell culture supernatant, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 30 pg/ml
  • Range

    24.58 pg/ml - 6000 pg/ml
  • Recovery

    94 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 96.43 84% - 104%
    Serum 94.53 82% - 103%
    Plasma 91.92 80% - 102%

  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Abcam’s TNF alpha Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human TNF alpha in serum, plasma and cell culture supernatants. (Human TNF alpha concentration is pretty low in normal serum/plasma, it may not be detected in this assay).

    This assay employs an antibody specific for Human TNF alpha coated on a 96-well plate. Standards and samples are pipetted into the wells and TNF alpha present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human TNF alpha antibody is added. After washing away unbound biotinylated antibody, HRP conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of TNF alpha bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

    Get higher sensitivity in only 90 minutes with Human TNF alpha ELISA Kit (ab181421) from our SimpleStep ELISA® range.


  • Notes

    Optimization may be required with urine samples.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform



  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    5X Assay Diluent B 1 x 15ml
    600X HRP-Streptavidin Concentrate 1 x 200µl
    Assay Diluent A 1 x 30ml
    Biotinylated anti-Human TNF alpha (lyophilized) 2 vials
    Recombinant Human TNF alpha Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
    TNF alpha Microplate (12 x 8 wells) 1 unit
  • Research areas

  • Function

    Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation.
  • Involvement in disease

    Genetic variations in TNF are a cause of susceptibility psoriatic arthritis (PSORAS) [MIM:607507]. PSORAS is an inflammatory, seronegative arthritis associated with psoriasis. It is a heterogeneous disorder ranging from a mild, non-destructive disease to a severe, progressive, erosive arthropathy. Five types of psoriatic arthritis have been defined: asymmetrical oligoarthritis characterized by primary involvement of the small joints of the fingers or toes; asymmetrical arthritis which involves the joints of the extremities; symmetrical polyarthritis characterized by a rheumatoidlike pattern that can involve hands, wrists, ankles, and feet; arthritis mutilans, which is a rare but deforming and destructive condition; arthritis of the sacroiliac joints and spine (psoriatic spondylitis).
  • Sequence similarities

    Belongs to the tumor necrosis factor family.
  • Post-translational

    The soluble form derives from the membrane form by proteolytic processing.
    The membrane form, but not the soluble form, is phosphorylated on serine residues. Dephosphorylation of the membrane form occurs by binding to soluble TNFRSF1A/TNFR1.
    O-glycosylated; glycans contain galactose, N-acetylgalactosamine and N-acetylneuraminic acid.
  • Cellular localization

    Secreted and Cell membrane.
  • Information by UniProt
  • Alternative names

    • APC1
    • APC1 protein
    • Cachectin
    • DIF
    • Differentiation inducing factor
    • Macrophage cytotoxic factor
    • Tnf
    • TNF superfamily member 2
    • TNF superfamily, member 2
    • TNF, macrophage derived
    • TNF, monocyte derived
    • TNF-a
    • TNF-alpha
    • TNFA
    • TNFSF2
    • Tumor necrosis factor
    • Tumor necrosis factor (TNF superfamily member 2)
    • Tumor necrosis factor alpha
    • Tumor necrosis factor ligand superfamily member 2
    • Tumor Necrosis Factor, Membrane Form
    • Tumor necrosis factor, soluble form
    see all
  • Database links

Associated products


Our Abpromise guarantee covers the use of ab100654 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.


  • Standard curve of huIL-8 with background signal subtracted (duplicates; +/- SD).
  • huTNFa measured in biological fluids, background signal subtracted (duplicates +/- SD).
  • TNFa detected in supernatants from control cells (C) or cells stimulated for 24 hours with 50 ng x mL-1 of PMA (ab120297) (P), and PMA with the addition of 1 ug x mL-1 of LPS (Sigma) (P+L) for the last 6 hours. PBMCs were stimulated for 48 hours with 2 % PHA-M (LifeTechnologies). Results shown after background signal was subtracted (duplicates +/- SD).
  • Representative standard curve using ab100654

  • Representative standard curve using ab100654



This product has been referenced in:

  • Okamgba OC  et al. Variations in the leukocyte and cytokine profiles between placental and maternal circulation in pregnancy-associated malaria. Res Rep Trop Med 9:1-8 (2018). Read more (PubMed: 30050350) »
  • Dada T  et al. Mindfulness Meditation Reduces Intraocular Pressure, Lowers Stress Biomarkers and Modulates Gene Expression in Glaucoma: A Randomized Controlled Trial. J Glaucoma 27:1061-1067 (2018). Read more (PubMed: 30256277) »
See all 5 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A


Thank you for contacting The Scientific Support team.    Since this kit was not specifically validated with urine (only serum, plasma, and cell culture supernatants) unfortunately we do not have concentration levels to report.  In addition, when we tested normal serum with this kit, we found the measured levels were below the sensitivity limit which is 30 pg/ml (and not 15pg/ml as specified in the protocol, which will soon be changed to fix this mistake).  Therefore if testing body fluids, I would recommend using this kit mainly for spiking and recovery studies or for testing diseased or treated samples which have expected elevated TNF-alpha levels than normal. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More


Thank you for contacting us. We do not validate our ELISA kits with monkey samples, but I was able to find that human TNF-alpha shares 94 – 95% amino acid sequence homology with monkey. We do have a human TNF-alpha ELISA kit ab100654, which may likely work with your samples.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More


Thank you for taking the time to provide more information.

I am sorry to hear you have had difficulty obtaining satisfactory results from this kit.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

Reviewing this case, I would like to offer some suggestions to help optimise the results from ab100654:

After reviewing the data I agree that the overall signals seem to be lower than expected, especially the standards. The most likely reason for this is that the Detection Antibody (Item F) was diluted with Assay Diluent A instead of Assay Diluent B.That's when we would see lower signals in tests.

I would also appreciate if you can confirm some further details:

What is the sample type?

At what temperature was the kit stored at and for how long?

Finally, this kit does not instruct the user to make a 666.7pg/ml or 74.07pg/ml standard.
The standard concentrations given in the kit are 6000, 2400, 960, 384, 153.6, 61.44, 24.58 and 0 pg/ml. Did you use a different dilution scheme?

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

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The equivalent IL10 assay, with 12 strips of 8 wells each, is ab100549. Product ab108871 comes with a single plate, not strips, but does use an HRP-conjugated streptavidin, as does the TNF alpha ELISA, ab100654.

Click here (or use the following: https://www.abcam.com/index.html?datasheet=100549).

Please contact us if you have any other questions.

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Thank you for your interest in our IL6 and TNF alpha ELISA kits. My colleague will be happy to discuss pricing with you. As I mentioned, she will ask how many kits you want of each type, IL6 and TNF alpha. If you have any technical questions, our Scientific Support team is available from 8:30 in the morning to 8:00 in the evenening, EST.  

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Thank you very much for contacting us. There is no fundamental difference between these kits. The ab46087 is based on sandwich ELISA procedure in which provided microtitre plate is coated with antibody. Ab100654 and ab108908 are based on direct ELISA procedure where antigens must be coated by incubating the plate with antigen or sample. We can assure that these KITS are equally suitable to detect TNF alpha in samples and are fully guaranteed by Abcam. I hope this information will be helpful. Please let us know if you have any questions or concerns.

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