Name: Human TNF alpha ELISA Kit
SKU: ab181421
Rating: 5 (4 reviews)

Human TNF alpha ELISA kit is a single-wash 90-min SimpleStep ELISA® used to quantify Human TNFα with a sensitivity of 4.32 pg/mL. The assay uses a simple Mix-Wash-Read protocol with just one incubation and wash step.

- Colorimetric assay - 450nm readout; works on any plate reader
- Different formats for different needs: 10x 96-well plates for bulk orders and 384-well for higher throughput
- Broad sample compatibility – works with CSF, serum, plasma, cell culture supernatants, and other biological fluids
- Cited in over 250 publications

Images

Sandwich ELISA - Human TNF alpha ELISA Kit (AB181421), expandable thumbnail

Publications

Key facts

Detection method: Colorimetric
Sample types: Cerebral Spinal Fluid, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
Assay type: Sandwich (quantitative)
Reactive species: Human
Range: 15.63 - 1000 pg/mL
Assay time: 1h 30m
Sensitivity: = 4.32 pg/mL

Abcam Recommends

Reactivity data

Application	Reactivity	Dilution info	Notes
Application sELISA	Reactivity Reacts	Dilution info -	Notes -

Associated Products

Select an associated product type

4 products for Alternative Product

Target data

See full target information TNF

Function

The TNF protein, primarily secreted by macrophages, binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It induces cell death in specific tumor cell lines and acts as a potent pyrogen, causing fever directly or by stimulating interleukin-1 secretion. TNF is implicated in cachexia induction and can stimulate cell proliferation and differentiation under certain conditions. It impairs regulatory T-cells (Treg) function in rheumatoid arthritis patients through FOXP3 dephosphorylation, upregulating protein phosphatase 1 (PP1), which dephosphorylates 'Ser-418' of FOXP3, inactivating FOXP3 and leading to defective Treg cells. TNF is a key mediator of cell death in the anticancer effect of BCG-stimulated neutrophils with DIABLO/SMAC mimetic in the RT4v6 bladder cancer cell line. It induces insulin resistance in adipocytes by inhibiting insulin-induced IRS1 tyrosine phosphorylation and glucose uptake. TNF plays a role in angiogenesis by inducing VEGF production with IL1B and IL6. Additionally, the TNF intracellular domain (ICD) form stimulates IL12 production in dendritic cells. This supplementary information is collated from multiple sources and compiled automatically.

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 384 Tests

Components

10X Wash Buffer PT (ab206977)

1 x 20 mL

384 well CaptSure™ microplates

1 x 1 Unit

Antibody Diluent 4BR

1 x 6 mL

Human TNFa Capture Antibody

1 x 600 µL

Human TNFa Detector Antibody

1 x 600 µL

Human TNFa Lyophilized Recombinant Protein

2 x 1 Vial

Plate Seals

1 x 1 Unit

Sample Diluent NS (ab193972)

2 x 50 mL

Stop Solution

2 x 12 mL

TMB Development Solution

2 x 12 mL

Key facts

Detection method: Colorimetric
Sample types: Cerebral Spinal Fluid, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
Assay type: Sandwich (quantitative)
Reactive species: Human
Range: 15.63 - 1000 pg/mL
Assay time: 1h 30m
Assay Platform: Microplate
Sensitivity: = 4.32 pg/mL

Precision

Intra assay

Sample	n	C.V.
Sample PBMC media	n 8	C.V. 2.5

Inter assay

Sample	n	C.V.
Sample PBMC media	n 3	C.V. 3.1

Recovery

Sample specific recovery

Sample type	Average %	Range
Sample type Cerebral Spinal Fluid	Average % = 83	Range 80 - 88 %
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

Human TNF alpha ELISA Kit ab181421 is a rapid single-wash 90-min ELISA kit to measure Human TNF alpha in cerebral spinal fluid, cell culture supernatant, serum and plasma. This SimpleStep ELISA® sensitivity is 4.32 pg/mL.

How the assay works

Human TNF alpha SimpleStep ELISA® 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.

Assay Specificity

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

Human TNF alpha ELISA Kit ab181421 protocol summary:

1. Mix: add samples/standards to the wells together with the capture and detector antibody cocktail. Incubate for 1 hr at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add stop solution
5. Read the results on a plate reader at 450nm

How other researchers are using Human TNF alpha ELISA Kit ab181421

Human TNF alpha ELISA Kit ab181421 has been used to assess inflammation in different physio-pathological contexts such as obesity⁽¹⁾, coronary artery disease⁽²⁾, and rheumatoid arthritis⁽³⁾.

References:

⁽¹⁾Shaohong Yu et al. 2023, PMID: 38156278,
⁽²⁾Yongshun Wang et al. 2023, PMID: 38054817,
⁽³⁾Yaju Tan et al. 2024, PMID: 37997635

Save your precious samples

Our 384-well Human TNF alpha ELISA Kit only requires a maxmum of 12.5µL

Design your own immunoassay

We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation

- Human/Monkey TNF alpha Antibody Pair - BSA and Azide free Human/Monkey TNF alpha Antibody Pair - BSA and Azide free ab241791
- Anti-TNF alpha antibody [EPR18565-29] - BSA and Azide free (Capture) Anti-TNF alpha antibody [EPR18565-29] - BSA and Azide free (Capture) ab242476
- Anti-TNF alpha antibody [EPR18565-33] - BSA and Azide free (Detector) Anti-TNF alpha antibody [EPR18565-33] - BSA and Azide free (Detector) ab242978

Related and recommended products

Human TNF alpha ELISA kit ab181421 is commonly used to measure TNF alpha as a marker of inflammation.

Other ELISA kits to measure inflammatory markers include:

- Human IL-6 ELISA kit Human IL-6 ELISA Kit ab178013
- Human IL-8 ELISA kit Human IL-8 ELISA Kit ab214030
- Human IL-1 beta ELISA kit Human IL-1 beta ELISA Kit ab214025

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

10 product images

Sandwich ELISA - Human TNF alpha ELISA Kit (ab181421)
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 values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human TNF alpha ELISA Kit (ab181421)
Human TNF alpha standard curve comparison.
Standard Curve comparison between human TNF alpha SimpleStep ELISA kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.
Sandwich ELISA - Human TNF alpha ELISA Kit (ab181421)
Interpolated concentrations of native TNF alpha in human PBMC cell culture supernatant samples.
PBMC cells were treated with 1.5% PHA-M for 46 hours in 10F RPMI. The concentrations of TNF alpha were measured in duplicates, interpolated from the TNF alpha standard curve and corrected for sample dilution. Undiluted samples are PHA-M treated PBMC cell culture supernatant 10%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TNF alpha concentration was determined to be 7326.31 pg/mL in PHA-M treated PBMC cell culture supernatant.
Sandwich ELISA - Human TNF alpha ELISA Kit (ab181421)
Interpolated concentrations of native TNF alpha PBMC cell culture supernatant samples.
PBMC cells were stimulated with 1.5% PHA-M or vehicle control in 10F RPMI and incubated for 46 hours. The concentrations of TNF alpha were measured in duplicate and interpolated from the TNF alpha standard curves. Undiluted samples are PHA-M stimulated PBMC cell culture supernatant 10% and unstimulated PBMC cell culture supernatant 10%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TNF alpha concentration was back calculated determined to be 7265.71 pg/mL in PHA-M stimulated PBMC cell culture supernatant and undetectable in the unstimulated PBMC control.
Sandwich ELISA - Human TNF alpha ELISA Kit (ab181421)
Linearity of dilution - native TNF alpha.
Native TNF alpha was measured in biological samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS.
Sandwich ELISA - Human TNF alpha ELISA Kit (ab181421)
Linearity of dilution - recombinant TNF alpha.
Recombinant TNF alpha was spiked biological samples and diluted in a 2-fold dilution series in Sample Diluent NS. Twenty individual healthy human female/male donors were measured in duplicate for the presence of TNF alpha. All values were below the detectable range of the assay
NL – Non-Linear
Sandwich ELISA - Human TNF alpha ELISA Kit (ab181421)
Human TNF alpha concentration was interpolated from the standard curve. Supernatants from cell culture samples were serially diluted and assessed by the Human TNF alpha ELISA kit (ab181421). Wild-type THP-1 and TNF alpha knockout THP-1 (Human TNF knockout THP-1 cell line ab273761) cells were assessed in duplicate (n=2). Cells were either treated with 100 ng/ml LPS for 16 h to induce expression of TNF alpha or not treated with LPS. Data are represented as the mean and error bars represent standard deviation."
Sandwich ELISA - Human TNF alpha ELISA Kit (ab181421)
Raw data value for example of TNF standard curve in Sample Diluent NS.
Raw data values are shown in the table
Sandwich ELISA - Human TNF alpha ELISA Kit (ab181421)
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
Sandwich ELISA - Human TNF alpha ELISA Kit (ab181421)
Example of human TNF alpha standard curve in 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed.