Human TNF alpha ELISA Kit

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Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 4.32 pg/ml
  • Range: 15.63 pg/ml – 1000 pg/ml
  • Sample type: Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

Overview

Product name

Human TNF alpha ELISA Kit
See all TNF alpha kits

Detection method

Colorimetric

Precision

Intra-assay
SamplenMeanSDCV%
PBMC media8  2.5%
Inter-assay
SamplenMeanSDCV%
PBMC media3  3.1%

Sample type

Cell culture supernatant, Serum, Hep Plasma, EDTA Plasma, Cit plasma

Assay type

Sandwich (quantitative)

Sensitivity

4.32 pg/ml

Range

15.63 pg/ml – 1000 pg/ml

Recovery

 

Sample specific recovery
Sample typeAverage %Range
Cell culture supernatant110105% – 113%
Serum8883% – 93%
Hep Plasma9889% – 110%
EDTA Plasma10095% – 104%
Cit plasma9186% – 97%

Assay time

1h 30m

Assay duration

One step assay

Species reactivity

Reacts with: Human

Product overview

Human TNF alpha ELISA kit (ab181421) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of TNF alpha protein in human serum, plasma and culture media. It uses our proprietary SimpleStep ELISA® technology. Quantitate human TNFa with 4.32 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 strips


A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpeStep ELISA® kits.


ASSAY SPECIFICITY This kit recognizes both native and recombinant human TNF alpha protein in serum, plasma, and cell culture supernatant samples only.


CROSS REACTIVITY Recombinant 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.

INTERFERENCE Recombinant Human TNF R1 was prepared at 1000 pg/mL and tested for interference. No interference with was observed.

SPECIES REACTIVITY This 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: Primate
Reactivity < 3% was determined for the following species: Mouse/Rat


CALIBRATION This 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).

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.

Platform

Microplate

Properties

Storage instructions

Store at +4°C. Please refer to protocols.
Components1 x 96 tests
10X Wash Buffer PT (ab206977)1 x 20ml
Antibody Diluent 4BR1 x 6ml
Human TNFa Capture Antibody1 x 600µl
Human TNFa Detector Antibody 1 x 600µl
Human TNFa Lyophilized Recombinant Protein2 vials
Plate Seals1 unit
Sample Diluent NS (ab193972)1 x 50ml
SimpleStep Pre-Coated 96-Well Microplate (ab206978)1 unit
Stop Solution1 x 12ml
TMB Development Solution1 x 12ml

Research areas

  • Immunology
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  • Innate Immunity
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  • Cytokines
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  • Necrosis Factors
  • Immunology
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  • Innate Immunity
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  • Cytokines
  •  
  • TNF Superfamily
  • Signal Transduction
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  • Growth Factors/Hormones
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  • TNF
  • Stem Cells
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  • Signaling Pathways
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  • TGF beta
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  • TNF alpha
  • Microbiology
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  • Organism
  •  
  • Virus
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  • RNA Virus
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  • ssRNA positive strand virus
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  • SARS Coronavirus
  • Cancer
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  • Growth factors
  •  
  • TNF
  • Cancer
  •  
  • Tumor immunology
  •  
  • Cytokines
  •  
  • TNF
  • Cardiovascular
  •  
  • Atherosclerosis
  •  
  • Vascular Inflammation
  •  
  • Inflammatory mediators
  • Kits/ Lysates/ Other
  •  
  • Kits
  •  
  • ELISA Kits
  •  
  • ELISA Kits
  •  
  • TNF ELISA kits
  • Metabolism
  •  
  • Types of disease
  •  
  • Metabolic disorders
  • Neuroscience
  •  
  • Development
  • Neuroscience
  •  
  • Diseases
  • Neuroscience
  •  
  • Processes

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
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 localization

Secreted and Cell membrane.

Protocols