• 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 protein
    • Cachectin
    • DIF
    • soluble form
    • Tnf
    • TNF a
    • TNF macrophage derived
    • TNF monocyte derived
    • TNF superfamily member 2
    • TNF-a
    • TNF-alpha
    • TNFSF2
    • Tumor necrosis factor
    • Tumor necrosis factor alpha
    • Tumor necrosis factor ligand superfamily member 2
    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:
  • Escribano-Lopez I  et al. The mitochondria-targeted antioxidant MitoQ modulates oxidative stress, inflammation and leukocyte-endothelium interactions in leukocytes isolated from type 2 diabetic patients. Redox Biol 10:200-205 (2016). WB ; Human . Read more (PubMed: 27810734) »

See 1 Publication for this product

Customer reviews and Q&As

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 tes...

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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 l...

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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 yo...

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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 follow...

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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 Scien...

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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 ELIS...

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