Mouse TNF alpha ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Mouse TNF alpha in Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma samples.
Images
Publications
- 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
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
Abcam Recommends
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
Associated Products
Select an associated product type
5 products for Alternative Product
1 products for Alternative Version
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).The TNF intracellular domain (ICD) form induces IL12 production in dendritic cells.
Alternative names
Tumor necrosis factor, Cachectin, TNF-alpha, Tumor necrosis factor ligand superfamily member 2, TNF-a, Tnfa, Tnf, Tnfsf2
What's included?
Recommended products
Mouse TNF alpha ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Mouse TNF alpha in Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma samples.
Alternative names
Tumor necrosis factor, Cachectin, TNF-alpha, Tumor necrosis factor ligand superfamily member 2, TNF-a, Tnfa, Tnf, Tnfsf2
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 | mean | SD | C.V. |
|---|---|---|---|---|
Sample Overall | n 8 | mean - | SD - | C.V. 6.7 |
Inter assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Overall | n 3 | mean - | SD - | 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 long-term storage conditions
+4°C
- Storage information
+4°C
Notes
Mouse TNF alpha ELISA Kit (ab208348) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of TNF alpha protein in cell culture supernatant, serum, cit plasma, edta plasma, and hep plasma. It uses our proprietary SimpleStep ELISA® technology. Quantitate Mouse TNF alpha with 9.1 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 (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
ASSAY SPECIFICITY
This kit recognizes both native and recombinant mouse TNF alpha protein in serum, plasma, and cell culture supernatant samples only.
CROSS REACTIVITY
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%.
INTERFERENCE
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.
SPECIES REACTIVITY
This kit recognizes mouse TNF alpha protein.
CALIBRATION
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)
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.
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.
14 product images
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 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 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 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed.
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.
Downloads
Product protocols
- Visit the general protocols
- Visit troubleshooting
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com