Range: 46.88 pg/ml - 3000 pg/ml Sample type: Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum Detection method: Colorimetric Assay type: Sandwich (quantitative) Reacts with: Mouse 产品概述 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 stripsA 384-well SimpleStep ELISA microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA kits.ASSAY SPECIFICITYThis kit recognizes both native and recombinant mouse TNF alpha protein in serum, plasma, and cell culture supernatant samples only.CROSS REACTIVITYRecombinant 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%.INTERFERENCERecombinant 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 REACTIVITYThis kit recognizes mouse TNF alpha protein.CALIBRATIONThis 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. 1 x 600µl10X Mouse TNF alpha Detector Antibody 1 x 600µl10X Wash Buffer PT (ab206977) 1 x 20mlAntibody DiluentCPI - HAMA Blocker (ab193969) 1 x 6mlMouse TNF alpha Lyophilized Recombinant Protein 2 vialsPlate Seals 1 unitSample Diluent NBS 1 x 20mlSample Diluent NS (ab193972) 1 x 50mlSimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unitStop Solution 1 x 12mlTMBDevelopment Solution 1 x 12ml 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. 疾病相关 Genetic variations in TNF are a cause of susceptibility psoriatic arthritis (PSORAS) [MIM:607507]. PSORAS is an inflammatory, seronegative arthritis associated with psoriasis. It is a heterogeneous disorder ranging from a mild, non-destructive disease to a severe, progressive, erosive arthropathy. Five types of psoriatic arthritis have been defined: asymmetrical oligoarthritis characterized by primary involvement of the small joints of the fingers or toes; asymmetrical arthritis which involves the joints of the extremities; symmetrical polyarthritis characterized by a rheumatoidlike pattern that can involve hands, wrists, ankles, and feet; arthritis mutilans, which is a rare but deforming and destructive condition; arthritis of the sacroiliac joints and spine (psoriatic spondylitis). 翻译后修饰 The soluble form derives from the membrane form by proteolytic processing.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.O-glycosylated; glycans contain galactose, N-acetylgalactosamine and N-acetylneuraminic acid. Tumor necrosis factor ligand superfamily member 2 Tumor Necrosis Factor, Membrane Form Tumor necrosis factor, soluble form see all 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. 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. 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. 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. 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). 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. 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 g/mL Kanamycin and 2 mM L-glutamine. Cells were starved for 24 hours and treated in the presence and absence of 5 g/mL of LPS. 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. To learn more about the advantages of recombinant antibodies see here. 发表研究结果有使用 ab208348？请让我们知道，以便我们可以引用本数据表中的参考文章。 ab208348 被引用在 55 文献中. Hou C et al. Mono-macrophage-Derived MANF Protects Against Lipopolysaccharide-Induced Acute Kidney Injury via Inhibiting Inflammation and Renal M1 Macrophages. Inflammation 44:693-703 (2021).PubMed: 33145627 Liu H et al. Catalytically potent and selective clusterzymes for modulation of neuroinflammation through single-atom substitutions. Nat Commun 12:114 (2021).PubMed: 33414464 Rao Y et al. Delivery of Long Non-coding RNA NEAT1 by Peripheral Blood Monouclear Cells-Derived Exosomes Promotes the Occurrence of Rheumatoid Arthritis via the MicroRNA-23a/MDM2/SIRT6 Axis. Front Cell Dev Biol 8:551681 (2020).PubMed: 33042992 Wang X et al. Phosphorylated STAT3 suppresses microRNA-19b/1281 to aggravate lung injury in mice with type 2 diabetes mellitus-associated pulmonary tuberculosis. J Cell Mol Med 24:13763-13774 (2020).PubMed: 33089914 Hu W et al. Blueberry anthocyanin-enriched extract ameliorates transverse aortic constriction-induced myocardial dysfunction via the DDAH1/ADMA/NO signaling pathway in mice. Mol Med Rep 21:454-462 (2020).PubMed: 31746378 View all Publications for this product The kit was used for mouse spleen lysates and with denervated muscle tissue. There is a very little response with lysates but it exists. With my experience, this kit works the best for cell culture supernatants. We ran this kit on mouse serum pools from C57BL6 mice and did not get any meaningful signal and it was off the curve. Do not run on serum of mice that are not chronically inflamed or are not treated with something that would lead to a spike in TNFa. Other than that, a good ELISA, standard curve looked good.Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES For licensing inquiries, please contact partnerships@abcam.com