OverviewProduct nameHuman TNF alpha ELISA KitSee all TNF alpha kitsDetection methodColorimetricPrecisionIntra-assaySamplenMeanSDCV%PBMC media82.5%Inter-assaySamplenMeanSDCV%PBMC media33.1%Sample typeCell culture supernatant, Serum, Hep Plasma, EDTA Plasma, Cit plasmaAssay typeSandwich (quantitative)Sensitivity 4.32 pg/mlRange15.63 pg/ml -1000 pg/mlRecoverySample specific recoverySample typeAverage %RangeCell culture supernatant110105%- 113%Serum8883%- 93%Hep Plasma9889%- 110%EDTA Plasma10095%- 104%Cit plasma9186%- 97%Assay time1h30mAssay durationOne step assaySpecies reactivityReacts with:HumanProduct overviewHuman TNF alpha ELISA kit (ab181421) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of TNF alpha protein inhuman serum, plasma and culture media. It uses our proprietary SimpleStep ELISA® technology. Quantitate human TNFa with4.32 pg/mL sensitivity.SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:-Single-wash protocol reduces assay time to 90 minutes or less-High sensitivity, specificity and reproducibility from superior antibodies-Fully validated in biological samples-96-wells plate breakable into 12 x 8 wells stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpeStep ELISA® kits.ASSAY SPECIFICITYThis kit recognizes both native and recombinant human TNF alpha protein in serum, plasma, and cell culture supernatant samples only.CROSS REACTIVITYRecombinant proteins were prepared at 1000 pg/mL and assayed for cross reactivity. No cross reactivity was found for the following targets: - Human IL-2 - Human IL-4 - Human IL-1 alpha - Human IFN gamma - Human TNF beta - Human TNF R1.INTERFERENCERecombinant Human TNF R1 was prepared at 1000 pg/mL and tested for interference. No interference with was observed.SPECIES REACTIVITYThis kit recognizes human TNF alpha protein. Other species reactivity was determined by measuring 1000 pg/mL recombinant proteins of various species, interpolating the protein concentrations from the human standard curve, and expressing the interpolated concentrations as a percentage of the protein concentration assayed at the same dilution. Reactive species: PrimateReactivity < 3%="" was="" determined="" for="" the="" following="" species:="">CALIBRATIONThis immunoassay is calibrated against a highly purified human TNF alpha. The NIBSC/WHO unclassified purified human TNF alpha preparation 12/154 was evaluated in this kit. The dose response curve of the unclassified standard TNF alpha parallels the SimpleStep standard curve. To convert sample values obtained with the SimpleStep Human TNF alpha kit to approximate NIBSC 12/154 International units, use the equation below.NIBSC (12/154) approximate value (IU/mL) = 0.094 x SimpleStep Human TNF alpha value (pg/mL).NotesTNF-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.PlatformMicroplatePropertiesStorage instructionsStore at +4°C. Please refer to protocols.Components1 x 96 tests10X Wash Buffer PT (ab206977)1 x 20mlAntibody Diluent 4BR1 x 6mlHuman TNFa Capture Antibody1 x 600µlHuman TNFa Detector Antibody1 x 600µlHuman TNFa Lyophilized Recombinant Protein2 vialsPlate Seals1 unitSample Diluent NS (ab193972)1 x 50mlSimpleStep Pre-Coated 96-Well Microplate (ab206978)1 unitStop Solution1 x 12mlTMBDevelopment Solution1 x 12mlResearch areasImmunologyInnate ImmunityCytokinesNecrosis FactorsImmunologyInnate ImmunityCytokinesTNF SuperfamilySignal TransductionGrowth Factors/HormonesTNFStem CellsSignaling PathwaysTGF betaTNF alphaCancerGrowth factorsTNFCancerTumor immunologyCytokinesTNFCardiovascularAtherosclerosisVascular InflammationInflammatory mediatorsKits/ Lysates/ OtherKitsELISA KitsELISA KitsTNF ELISA kitsMetabolismTypes of diseaseMetabolic disordersFunctionCytokine 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-translationalmodificationsThe 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.Target information above from: UniProt accessionP01375The UniProt ConsortiumThe Universal Protein Resource (UniProt) in 2010Nucleic Acids Res. 38:D142-D148 (2010).Information by UniProtAlternative namesAPC1APC1 proteinCachectinDIFDifferentiation inducing factorMacrophage cytotoxic factorTnfTNF superfamily member 2TNF superfamily, member 2TNF, macrophage derivedTNF, monocyte derivedTNF-aTNF-alphaTNFATNFA_HUMANTNFSF2Tumor necrosis factorTumor necrosis factor (TNF superfamily member 2)Tumor necrosis factor alphaTumor necrosis factor ligand superfamily member 2Tumor Necrosis Factor, Membrane FormTumor necrosis factor, soluble formsee allDatabase linksEntrez Gene:7124 HumanOmim:191160 HumanSwissProt:P01375 HumanUnigene:241570 Human