Overview

  • Product nameHuman TNF alpha ELISA Kit
    See all TNF alpha kits
  • Detection methodColorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    PBMC media 8 2.3%
    Inter-assay
    Sample n Mean SD CV%
    PBMC media 3 5%
  • Sample type
    Serum, Plasma, Cell culture media
  • Assay typeSandwich (quantitative)
  • Sensitivity
    14 pg/ml
  • Range
    31.25 pg/ml - 2000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 99.5 80% - 119%
    Cell culture media 100.97 95% - 109%
    Heparin Plasma 98 91% - 106%
    EDTA Plasma 102 95.7% - 110%
    Citrate Plasma 93.09 83.5% - 99%

  • Assay time
    1h 30m
  • Assay durationOne step assay
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse
  • Product overview

    Abcam’s TNF-alpha in vitro SimpleStep ELISA®  (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of TNF-alpha protein in Human serum, plasma and culture media.

    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    TNF-alpha, also known as cachectin or TNFSF1A, is the prototypic ligand of the TNF superfamily which plays a central role in inflammation, apoptosis, proliferation, invasion, angiogenesis, metastasis and morphogenesis. It is expressed on macrophages, endothelial, epithelial and tumor cells as a 26kDa transmembrane protein. TNF-alpha is cleaved by proteolytic processing into six chains: (1) TNF membrane form, (2) Intracellular domain 1, (3) Intracellular domain 2, (4) C-domain 1, (5) C-domain 2 and (6) TNF soluble form. Signaling from TNF-alpha differs depending on the type of ligand initiating the signaling event (intracellular, membrane or soluble). As an example, the membrane form of TNF-alpha appears to mediate anti-tumorigenic therapeutic responses whereas the soluble ligand is linked to inflammation and proliferation.

  • Tested applicationsSuitable for: Sandwich ELISAmore details
  • PlatformMicroplate

Properties

  • FunctionCytokine 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 diseaseGenetic 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 similaritiesBelongs to the tumor necrosis factor family.
  • Post-translational
    modifications
    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 localizationSecreted 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
    • TNFA_HUMAN
    • TNFSF2
    • Tumor necrosis factor
    • Tumor necrosis factor alpha
    • Tumor necrosis factor ligand superfamily member 2
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab181421 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.

Human TNF alpha ELISA Kit images

  • Background-subtracted data values (mean +/- SD) are graphed. 

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Human PBMCs were cultured in RPMI supplemented with 10% fetal calf serum, 2mM L-glutamine, 100 U/mL penicillin and 100 µg/mL streptomycin.  Cells were cultured for 2 days at 37°C in the presence or absence of PHA.  The concentrations of TNF-alpha were interpolated from the calibration curve and corrected for sample dilution.  The mean TNF-alpha concentration was determined to be undetectable in basal media, 1.2 ng/mL in unstimulated PMBC supernatants and 4.7 ng/mL in stimulated PBMC supernatants. 

Protocols

References for Human TNF alpha ELISA Kit (ab181421)

This product has been referenced in:
  • Murphy RA  et al. Establishment of HK-2 Cells as a Relevant Model to Study Tenofovir-Induced Cytotoxicity. Int J Mol Sci 18:N/A (2017). ELISA ; Human . Read more (PubMed: 28257038) »
  • Zhou Y  et al. Enhancement of CD3AK cell proliferation and killing ability by a-Thujone. Int Immunopharmacol 30:57-61 (2016). Sandwich ELISA ; Human . Read more (PubMed: 26655741) »

See all 3 Publications for this product

Product Wall

efficient and accurate

Excellent Excellent 5/5 (Ease of Use)
Abreviews
I used this kit to detect TNF-a in stimulated human CD14+ monocytes. The assay was extremely efficient. There is no overnight coating step, and detection/capture antibodies are added as a cocktail at the same time as the samples. The assay was quite accurate, detecting even minimal amounts of TNF-a. I would use this assay again for its efficiency, requiring very little time at the bench.
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Verified customer

Submitted Jun 01 2016

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"