Mouse TNF alpha ELISA kit is a single-wash, 90-minute SimpleStep ELISA® designed to quantify mouse TNF alpha with a sensitivity of 9.1 pg/mL. The assay uses a simple Mix-Wash-Read protocol with just one incubation and wash step.
- Colorimetric sandwich ELISA -450 nm readout; works on any plate reader
- Different formats for different needs – 384-well format for higher throughput
- Broad sample compatibility – works with serum, plasma (heparin, citrate, EDTA), and cell culture supernatants
- Cited in over 235 publications
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- Biomolecules & biomedicine 24:505-5192024Silencing METTL14 alleviates liver injury in non-alcoholic fatty liver disease by regulating mitochondrial homeostasis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesWei Wang,Jun Yan,Long Han,Zi-Lin Zou,Ai-Lei XuPubMed 37902450
- NPJ systems biology and applications 10:52024The antidepressive mechanism of Longya Lilium combined with Fluoxetine in mice with depression-like behaviors.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHuina Ma,Hehua Huang,Chenyu Li,Shasha Li,Juefang Gan,Chunrong Lian,Yanwu LingPubMed 38218856
- Scientific reports 13:224092023Chia seeds oil ameliorate chronic immobilization stress-induced neurodisturbance in rat brains via activation of the antioxidant/anti-inflammatory/antiapoptotic signaling pathways.Applications:Unspecified applicationReactive species:Unspecified reactive speciesNorhan E Khalifa,Ahmed E Noreldin,Asmaa F Khafaga,Mohamed El-Beskawy,Eman Khalifa,Ali H El-Far,Abdel-Hasseb A Fayed,Abdeldayem ZakariaPubMed 38104182
- The European journal of neuroscience 59:36-532023Dexmedetomidine alleviates hippocampal neuronal loss and cognitive decline in rats undergoing open surgery under sevoflurane anaesthesia by suppressing CCAAT/enhancer-binding protein beta.Applications:Unspecified applicationReactive species:Unspecified reactive speciesShanshan Fu,Xianghai Zhao,Yingna Li,Xinwen Fan,Zeqing HuangPubMed 37985440
- Journal of inflammation research 16:5095-51092023LPA Alleviates Septic Acute Lung Injury via Protective Endothelial Barrier Function Through Activation of PLC-PKC-FAK.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRuifeng Bai,Jianqiu Pei,Shengqiang Pei,Xiangfeng Cong,Jerold Chun,Fang Wang,Xi ChenPubMed 38026263
- Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology 18:628-6392023GPNMB Ameliorates Neuroinflammation Via the Modulation of AMPK/NFκB Signaling Pathway After SAH in Mice.Applications:Unspecified applicationReactive species:Unspecified reactive speciesTao Li,Yuansheng Zhang,Qixiong Lu,Li Lei,Jingshu Du,Xiaoyang LuPubMed 37919457
- Parasitology research 122:3213-32312023Effects of metformin on parasitological, pathological changes in the brain and liver and immunological aspects during visceral toxocariasis in mice.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAmina M Salama,Rasha A Elmahy,Hoda A Ibrahim,Alaa Ibrahim Mohamed Amer,Asmaa Fawzy Eltantawy,Dina I ElgendyPubMed 37874393
- Biomedical reports 19:1002023Dexmedetomidine attenuates the neuroinflammation and cognitive dysfunction in aged mice by targeting the SNHG14/miR‑340/NF‑κB axis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesGuangbao He,Hongwei Ni,Kai Wang,Hongmei Gao,Yu Li,Jiaqiu Gu,Xin Ni,Zhiming Wang,Yang BaoPubMed 37954634
- Molecular medicine (Cambridge, Mass.) 29:1372023Resveratrol reduces ROS-induced ferroptosis by activating SIRT3 and compensating the GSH/GPX4 pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesXingjie Wang,Tianli Shen,Jie Lian,Kai Deng,Chao Qu,Enmeng Li,Gan Li,Yiwei Ren,Zijun Wang,Zhengdong Jiang,Xuejun Sun,Xuqi LiPubMed 37858064
- Heliyon 9:e201242023IL-1β-pretreated bone mesenchymal stem cell-derived exosomes alleviate septic endoplasmic reticulum stress via regulating SIRT1/ERK pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesXinsheng Cheng,Shikai Wang,Zhipeng Li,Di He,Jianguo Wu,Weiwei DingPubMed 37771539
Key facts
- Detection method
- Colorimetric
- Sample types
- Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Mouse
- Range
- 46.88 - 3000 pg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 9.1 pg/mL
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Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
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 (By similarity). 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 (PubMed:25586176). Plays a role in angiogenesis by inducing VEGF production synergistically with IL1B and IL6 (By similarity). Promotes osteoclastogenesis and therefore mediates bone resorption (PubMed:32741026). 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?
Recommended products
Mouse TNF alpha ELISA kit is a single-wash, 90-minute SimpleStep ELISA® designed to quantify mouse TNF alpha with a sensitivity of 9.1 pg/mL. The assay uses a simple Mix-Wash-Read protocol with just one incubation and wash step.
- Colorimetric sandwich ELISA -450 nm readout; works on any plate reader
- Different formats for different needs – 384-well format for higher throughput
- Broad sample compatibility – works with serum, plasma (heparin, citrate, EDTA), and cell culture supernatants
- Cited in over 235 publications
Key facts
- Detection method
- Colorimetric
- Sample types
- Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Mouse
- Range
- 46.88 - 3000 pg/mL
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 9.1 pg/mL
Precision
Intra assay
| Sample | n | C.V. |
|---|---|---|
Sample Overall | n 8 | C.V. 6.7 |
Inter assay
| Sample | n | C.V. |
|---|---|---|
Sample Overall | n 3 | C.V. 9.8 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Serum | Average % = 97 | Range 85 - 115 % |
Sample type EDTA Plasma | Average % = 99 | Range 98 - 101 % |
Sample type Heparin Plasma | Average % = 92 | Range 85 - 103 % |
Sample type Citrate plasma | Average % = 87 | Range 85 - 88 % |
Sample type Cell culture media | Average % = 111 | Range 109 - 115 % |
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
Mouse TNF alpha ELISA Kit ab208348 is a rapid single-wash 90-min ELISA kit to measure cell culture supernatant, Serum, plasma. This SimpleStep ELISA® sensitivity is 9.1 pg/mL.
How the assay works
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 mouse TNF alpha protein in serum, plasma, and cell culture supernatant samples only.
Recombinant mouse TNF RI, TNF RII, OPG, CD40 receptor, IL1 beta, IL-16, and IL-5 were prepared at 100 ng/mL and assayed for cross reactivity. No cross-reactivity was observed. Recombinant human TNF alpha was prepared at 100 ng/mL, 25 ng/mL, and 3 ng/mL and assayed for cross reactivity. On average, 3.0% cross-reactivity was observed with a standard deviation of 0.7%.
Recombinant mouse TNF RI, TNF RII, OPG, CD40 receptor, IL1 beta, IL-16, and IL-5 and recombinant human TNF alpha were prepared at 100 ng/mL and tested for interference. No interference with was observed.
This immunoassay is calibrated against a highly purified mouse TNF alpha. The NIBSC/WHO unclassified purified mouse TNF alpha preparation 88/532 was evaluated in this kit. The dose response curve of the unclassified standard 88/532 parallels the SimpleStep standard curve. To convert sample values obtained with the SimpleStep mouse TNF alpha kit to approximate NIBSC 88/532 units, use the equation below.
NIBSC (88/532) approximate value (units/mL) = 2.0 x SimpleStep mouse TNF alpha value (pg/mL)
Mouse TNF alpha ELISA Kit ab208348 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 Mouse TNF alpha ELISA Kit ab208348
Mouse TNF alpha ELISA Kit ab208348 has been used to assess inflammation in different physio-pathological contexts such as liver disease(1), aging(2), and Alzheimer's disease(3).
References:
(1)Ai-Lei Xu et al. 2023, PMID: 37902450,
(2)Yang Bao et al. 2023, PMID: 37954634,
(3)Ramon Velazquez et al. 2023, PMID: 37548694
Save your precious samples
Our 384-well Mouse 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
- Mouse TNF alpha Antibody Pair - BSA and Azide free Mouse TNF alpha Antibody Pair - BSA and Azide free ab241672
- Anti-TNF alpha antibody [EPR16803-2] - BSA and Azide free (Capture) Anti-TNF alpha antibody [EPR16803-2] - BSA and Azide free (Capture) ab242949
- Anti-TNF alpha antibody [EPR16803-84] - BSA and Azide free (Detector) Anti-TNF alpha antibody [EPR16803-84] - BSA and Azide free (Detector) ab242977
Related and recommended products
Mouse TNF alpha ELISA kit ab208348 is commonly used to measure TNF alpha as a marker of inflammation.
Other ELISA kits to measure inflammatory markers include:
- Mouse IL-6 ELISA kit Mouse IL-6 ELISA Kit ab222503
- Mouse IFN gamma ELISA kit Mouse IFN gamma ELISA Kit ab282874
- Mouse IL-1 beta ELISA kit Mouse IL-1 beta ELISA Kit ab197742
Tumor necrosis factor, or TNF alpha, is a cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. Furthermore, TNF alpha is a ligand of the TNF superfamily which plays a central role in inflammation, apoptosis, proliferation, invasion, angiogenesis, metastasis, and morphogenesis. It is mainly secreted by macrophages, and can induce cell death of certain tumor cell lines. Rat and Human TNF alpha are 95% and 79% identical to Mouse TNF alpha, respectively. TNF alpha is expressed on macrophages and endothelial, epithelial, and tumor cells as a 26 kDa transmembrane protein. TNF-alpha is cleaved by proteolytic processing into six chains: (1) TNF membrane form, (2) Intracellular domain 1 (ICD1), (3) Intracellular domain 2 (ICD2), (4) C-domain 1, (5) C-domain 2 and (6) TNF soluble form. ICD1 and ICD2 are released into the cytosol, while C-domain 1 and C-domain 2 are released into the extracellular space. Furthermore, 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. 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. Likewise, the TNF intracellular domain (ICD) form induces IL-12 production in dendritic cells.
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14 product images
Sandwich ELISA - Mouse TNF alpha ELISA Kit (ab208348)
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 - Mouse TNF alpha ELISA Kit (ab208348)
Example of mouse TNF alpha standard curve in Sample Diluent 25BS.
Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Mouse TNF alpha ELISA Kit (ab208348)
Example of mouse TNF alpha standard curve in Sample Diluent 25BS.
The TNF alpha standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Mouse TNF alpha ELISA Kit (ab208348)
Example of mouse TNF alpha standard curve in Sample Diluent 10BS.
The TNF alpha standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Mouse TNF alpha ELISA Kit (ab208348)
Example of mouse TNF alpha standard curve in Sample Diluent NS.
Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Mouse TNF alpha ELISA Kit (ab208348)
Example of mouse TNF alpha standard curve in Sample Diluent NS.
The TNF alpha standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Mouse TNF alpha ELISA Kit (ab208348)
Interpolated concentrations of spiked TNF alpha in mouse plasma samples.
The concentrations of TNF alpha were measured in duplicates, interpolated from the TNF alpha standard curves and corrected for sample dilution. Undiluted samples are as follows: plasma (citrate) 100%, plasma (EDTA) 100%, and plasma (heparin) 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TNF alpha concentration was determined to be 1.68 ng/mL in plasma (citrate), 1.81 ng/mL in plasma (EDTA) and 1.47 ng/mL in plasma (heparin).
Sandwich ELISA - Mouse TNF alpha ELISA Kit (ab208348)
Example of mouse TNF alpha standard curve in Sample Diluent 10BS.
Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Mouse TNF alpha ELISA Kit (ab208348)
Interpolated concentrations of spiked mouse TNF alpha in mouse serum and cell culture samples and native mouse TNF alpha in mouse RAW 264.7 LPS stimulated cell culture supernatants.
The concentrations of TNF alpha were measured in duplicates, interpolated from the TNF alpha standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 100%, cell culture 100%, and RAW 264.7 LPS stimulated cell culture supernatants 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TNF alpha concentration was determined to be 1.63 ng/mL in serum, 1.00 ng/mL in cell culture and 6.07 ng/mL in RAW 264.7 LPS stimulated cell culture supernatants.
Sandwich ELISA - Mouse TNF alpha ELISA Kit (ab208348)
Interpolated concentrations of native TNF alpha in mouse RAW 264.7 LPS stimulated and unstimulated cell culture supernatant samples.
The concentrations of TNF alpha were measured in duplicates, interpolated from the TNF alpha standard curves and corrected for sample dilution. Undiluted samples are as follows: RAW 264.7 LPS stimulated supernatant 50% and RAW 264.7 LPS unstimulated supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TNF alpha concentration was determined to be 6346.4 pg/mL in RAW 264.7 LPS stimulated supernatant and 83.7 pg/mL in RAW 264.7 LPS unstimulated supernatant. RAW 264.7 cells were cultured in HGDMEM with 100 μg/mL Kanamycin and 2 mM L-glutamine. Cells were starved for 24 hours and treated in the presence and absence of 5 μg/mL of LPS.
Sandwich ELISA - Mouse TNF alpha ELISA Kit (ab208348)
Assay sensitivity.
The calculated minimal detectable dose (MDD) is determined by calculating the mean of zero standard replicates and adding 2 standard deviations then extrapolating the corresponding concentrations. The MDD is dependent on the Sample Diluent buffer used.
Sandwich ELISA - Mouse TNF alpha ELISA Kit (ab208348)
Example of mouse TNF alpha standard curve in Sample Diluent 25BS in 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Mouse TNF alpha ELISA Kit (ab208348)
Example of mouse 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 - Mouse TNF alpha ELISA Kit (ab208348)
Example of mouse TNF alpha standard curve in Sample Diluent 10BS in 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed.
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