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AB181421

Human TNF alpha ELISA Kit

5

(4 Reviews)

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(257 Publications)

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

View Alternative Names

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

10 Images
Sandwich ELISA - Human TNF alpha ELISA Kit (AB181421)
  • sELISA

Supplier Data

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)
  • sELISA

Lab

Sandwich ELISA - Human TNF alpha ELISA Kit (AB181421)

Example of human TNF alpha standard curve in Sample Diluent NS in 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human TNF alpha ELISA Kit (AB181421)
  • sELISA

Unknown

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)
  • sELISA

Supplier Data

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)
  • sELISA

Lab

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 (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)
  • sELISA

Unknown

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)
  • sELISA

Unknown

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)
  • sELISA

Supplier Data

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)
  • sELISA

Supplier Data

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)
  • sELISA

Lab

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.

Key facts

Detection method

Colorimetric

Sample types

Cerebral Spinal Fluid, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma

Reacts with

Human

Assay type

Sandwich (quantitative)

Sensitivity

= 4.32 pg/mL

Range

15.63 - 1000 pg/mL

Assay time

1h 30m

Assay Platform

Microplate

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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You may be interested in:

AB241791

Human/Monkey TNF alpha Antibody Pair - BSA and Azide free

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We recommend this product because it’s often used in the same experiment or related research.

We advise that you always check the datasheet to ensure it fits your experiments, or contact ourtechnical teamfor help.

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Product details

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 ab241791
- Anti-TNF alpha antibody [EPR18565-29] - BSA and Azide free (Capture) ab242476
- 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 ab178013
- Human IL-8 ELISA kit ab214030
- 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.

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Precision

[ { "reproducibilityType": "Inter", "sample": "PBMC media", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "3.1" }, { "reproducibilityType": "Intra", "sample": "PBMC media", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "2.5" } ]
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Recovery

[ { "sample": "Cerebral Spinal Fluid", "range": "80 - 88 %", "average": "= 83" }, { "sample": "Cell culture supernatant", "range": "105 - 113 %", "average": "= 110" }, { "sample": "Serum", "range": "83 - 93 %", "average": "= 88" }, { "sample": "EDTA Plasma", "range": "95 - 104 %", "average": "= 100" }, { "sample": "Heparin Plasma", "range": "89 - 110 %", "average": "= 98" }, { "sample": "Citrate plasma", "range": "86 - 97 %", "average": "= 91" } ]
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What's included?

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Properties and storage information

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

This supplementary information is collated from multiple sources and compiled automatically.

Tumor necrosis factor alpha (TNF alpha) also known as TNF-a is a cytokine with a molecular weight of approximately 17 kDa. This protein is expressed mainly by macrophages though other cells like T-cells and natural killer cells also produce it. TNF alpha helps in regulating immune responses and inflammatory reactions. Its expression is important in triggering cell apoptosis and modulating immune cell activities. TNF alpha exists in membrane-bound and soluble forms both having biologic activity.
Biological function summary

TNF alpha acts as an important regulator of inflammatory and immune responses. It is not part of a complex but interacts with TNF receptors to activate various signaling cascades. TNF alpha promotes leukocyte recruitment to sites of infection or injury by enhancing adhesion molecule expression on endothelial cells. Additionally it induces the production of other pro-inflammatory cytokines like IL-6 and IL-1 amplifying the immune response. Researchers often measure TNF alpha levels in samples using methods like ELISA or enzyme-linked immunosorbent assay and develop specific antibodies like anti-TNF alpha for experimental and therapeutic purposes.

Pathways

TNF alpha participates in the NF-kB and MAPK pathways significant for mediating inflammatory and immune responses. In the NF-kB pathway TNF alpha activates transcription factors that control the expression of genes involved in inflammation and cell survival. TNF alpha also interacts with proteins such as the TNF receptor 1 (TNFR1) and receptor-associated factors (TRAFs) further relaying signals within these pathways. There is extensive research employing animal models using mouse TNF-alpha and anti-TNF reagents to dissect these pathways’ intricacies.

TNF alpha substantially affects conditions like rheumatoid arthritis and Crohn's disease both characterized by chronic inflammation. Elevated levels of TNF alpha drive the inflammatory processes central to these diseases leading to tissue damage. Other proteins like IL-6 also play critical roles in these disorders and together with TNF alpha create a pro-inflammatory environment. Therapeutic agents such as anti-TNF treatments aim to mitigate these effects by specifically targeting TNF alpha activity.
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Product protocols

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

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.
See full target information TNF
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Publications (257)

Recent publications for all applications. Explore the full list and refine your search

Frontiers in nutrition 10:1270435 PubMed38156278

2024

Serum levels of Vanin-2 increase with obesity in relation to inflammation of adipose tissue and may be a predictor of bariatric surgery outcomes.

Applications

Unspecified application

Species

Unspecified reactive species

Shan Geng,Dongmei Chen,Yanping Wang,Xingrui Yu,Dan Zuo,Xinlu Lv,Xuelian Zhou,Chengju Hu,Xuesong Yang,Xujue Ma,Wenjing Hu,Jiazhuang Xi,Shaohong Yu

Aging 15:14720-14732 PubMed38054817

2023

Meteorin-like protein elevation post-exercise improved vascular inflammation among coronary artery disease patients by downregulating NLRP3 inflammasome activity.

Applications

Unspecified application

Species

Unspecified reactive species

Jingjin Liu,Liwei Diao,Weiyi Xia,Xiaoyi Zeng,Wen Li,Jieru Zou,Tiansheng Liu,Xinli Pang,Yongshun Wang

International journal of rheumatic diseases 27:e14976 PubMed37997635

2023

Forsythiaside A exhibits anti-migration and anti-inflammation effects in rheumatoid arthritis in vitro model.

Applications

Unspecified application

Species

Unspecified reactive species

Minhui Su,Dinghua Zhou,Jiamin Huang,Ting Yang,Qi Zhou,Yaju Tan

Nature communications 14:7639 PubMed37993431

2023

Acoustofluidic Interfaces for the Mechanobiological Secretome of MSCs.

Applications

Unspecified application

Species

Unspecified reactive species

Ye He,Shujie Yang,Pengzhan Liu,Ke Li,Ke Jin,Ryan Becker,Jinxin Zhang,Chuanchuan Lin,Jianping Xia,Zhehan Ma,Zhiteng Ma,Ruoyu Zhong,Luke P Lee,Tony Jun Huang

Experimental and therapeutic medicine 26:575 PubMed38023363

2023

IGF2BP2‑dependent STIM1 inhibition protects against LPS‑induced pneumonia by alleviating endoplasmic reticulum stress and the inflammatory response.

Applications

Unspecified application

Species

Unspecified reactive species

Wei Zhou,Qigang Dai,Ning Su,Zhihui Liu,Jinxing Hu

Scientific reports 13:17344 PubMed37833265

2023

Increased blood immune regulatory cells in severe COVID-19 with autoantibodies to type I interferons.

Applications

Unspecified application

Species

Unspecified reactive species

Fatemeh Saheb Sharif-Askari,Narjes Saheb Sharif-Askari,Shirin Hafezi,Hawra Ali Hussain Alsayed,Balachandar Selvakumar,Mariam Wed Abdelaziz Eladham,Bushra Mdkhana,Ola Salam Bayram,Mohamad-Hani Temsah,Rabih Halwani

Heliyon 9:e20771 PubMed37842584

2023

Electroacupuncture combined with extracorporeal shock wave therapy improves pain symptoms and inflammatory factor levels in knee osteoarthritis patients.

Applications

Unspecified application

Species

Unspecified reactive species

Shengfu Liu,Qiudan Chen,Qinggang Zhang,Kun Tao,Changhong Li,Baolei Chang,Weifeng Wang,Zhong Wu

AMB Express 13:107 PubMed37789186

2023

Bioactive metabolites identified from Aspergillus terreus derived from soil.

Applications

Unspecified application

Species

Unspecified reactive species

Menna Fayek,Hassan Y Ebrahim,Mohamed S Abdel-Aziz,Heba Taha,Fatma A Moharram

Molecular medicine reports 28: PubMed37772394

2023

Kynurenic acid inhibits macrophage pyroptosis by suppressing ROS production via activation of the NRF2 pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Yuwei Gao,Xiaohui Guo,Yunpeng Zhou,Jie Du,Chengbo Lu,Lei Zhang,Siyuan Sun,Shengfang Wang,Yang Li

In vivo (Athens, Greece) 37:2028-2038 PubMed37652511

2023

Therapeutic Effect of Human Adipocyte-derived Stem Cell-derived Exosomes on a Transgenic Mouse Model of Parkinson's Disease.

Applications

Unspecified application

Species

Unspecified reactive species

Lung Chan,Wayne Hsu,Kai-Yun Chen,Weu Wang,Yi-Chieh Hung,Chien-Tai Hong
View all publications
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions
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