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Human TNF alpha ELISA Kit, Fluorescent is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human TNF alpha , Fluorescent in Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma samples.

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Sandwich ELISA - Human TNF alpha ELISA Kit, Fluorescent (AB229399), expandable thumbnail

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Key facts

Detection method
Fluorescent
Sample types
Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
Assay type
Sandwich (quantitative)
Reactive species
Human
Range
1.95 - 8000 pg/mL
Assay time
1h 30m
Sensitivity
= 1.23 pg/mL

Reactivity data

Application
sELISA
Reactivity
Reacts
Dilution info
-
Notes

-

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Target data

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. Impairs regulatory T-cells (Treg) function in individuals with rheumatoid arthritis via FOXP3 dephosphorylation. Up-regulates the expression of protein phosphatase 1 (PP1), which dephosphorylates the key 'Ser-418' residue of FOXP3, thereby inactivating FOXP3 and rendering Treg cells functionally defective (PubMed:23396208). Key mediator of cell death in the anticancer action of BCG-stimulated neutrophils in combination with DIABLO/SMAC mimetic in the RT4v6 bladder cancer cell line (PubMed:16829952, PubMed:22517918, PubMed:23396208). Induces insulin resistance in adipocytes via inhibition of insulin-induced IRS1 tyrosine phosphorylation and insulin-induced glucose uptake. Induces GKAP42 protein degradation in adipocytes which is partially responsible for TNF-induced insulin resistance (By similarity). Plays a role in angiogenesis by inducing VEGF production synergistically with IL1B and IL6 (PubMed:12794819). Promotes osteoclastogenesis and therefore mediates bone resorption (By similarity). The TNF intracellular domain (ICD) form induces IL12 production in dendritic cells.

Alternative names

TNFA, TNFSF2, TNF, Tumor necrosis factor, Cachectin, TNF-alpha, Tumor necrosis factor ligand superfamily member 2, TNF-a

What's included?

1 x 96 Tests
Components
100X Stoplight Red Substrate
1 x 120 µL
10X Wash Buffer PT (ab206977)
1 x 20 mL
500X Hydrogen Peroxide (H2O2, 3%)
1 x 50 µL
Antibody Diluent 5BI
1 x 6 mL
Human TNFa Capture Antibody
2 x 1 Vial
Human TNFa Detector Antibody
2 x 1 Vial
Human TNFa Lyophilized Recombinant Protein
2 x 1 Vial
Plate Seals
1 x 1 Unit
Sample Diluent 50BP
1 x 20 mL
Sample Diluent NS (ab193972)
1 x 50 mL
SimpleStep Pre-Coated Black 96-Well Microplate
1 x 1 Unit
Stoplight Red Substrate Buffer
1 x 12 mL

Recommended products

Human TNF alpha ELISA Kit, Fluorescent is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human TNF alpha , Fluorescent in Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma samples.

Key facts

Detection method
Fluorescent
Sample types
Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
Assay type
Sandwich (quantitative)
Reactive species
Human
Range
1.95 - 8000 pg/mL
Assay time
1h 30m
Assay Platform
Pre-coated microplate (12 x 8 well strips)
Sensitivity
= 1.23 pg/mL

Precision

Intra assay

Sample
Supernatant
n
8
mean
-
SD
-
C.V.
2.5

Inter assay

Sample
Supernatant
n
3
mean
-
SD
-
C.V.
3.1

Recovery

Sample specific recovery

Sample type
Cell culture supernatant
Average %
= 110
Range
105 - 113 %
Sample type
Serum
Average %
= 88
Range
83 - 93 %
Sample type
EDTA Plasma
Average %
= 100
Range
95 - 104 %
Sample type
Heparin Plasma
Average %
= 98
Range
89 - 110 %
Sample type
Citrate plasma
Average %
= 91
Range
86 - 97 %

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C

Notes

TNF-alpha in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of TNF-alpha protein in human serum, plasma, and cell culture supernatants.

This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
If using a Molecular Devices' plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.

The CatchPoint 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. CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.

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.

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1 product image

  • Sandwich ELISA - Human TNF alpha ELISA Kit, Fluorescent (ab229399), expandable thumbnail

    Sandwich ELISA - Human TNF alpha ELISA Kit, Fluorescent (ab229399)

    Example of human TNF alpha standard curve in Sample Diluent NS.

    The TNF alpha standard curve was prepared as described in Section 10. Raw data generated on SpectraMax M4 Multi-Mode Microplate Reader is shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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