Rat TNF-alpha ELISA Kit (Colorimetric)

ELISA: TNF-alpha ELISA Kit [Biotin] [NBP1-92681] - Standard curve

• Reactivity: Rt
• Applications: ELISA
• Suitable Sample Type: Cell culture supernatant, serum
• Standard Curve Range: 39.1 - 2,500 pg/mL
• Sensitivity: 11.0 pg/mL

Rat TNF-alpha ELISA Kit (Colorimetric) Summary

• Description: The rat TNF-alpha ELISA is an enzyme-linked immunosorbent assay for the quantitative detection of rat TNF-alpha.
• Specificity: The interference of circulating factors of the immune systemwas evaluated by spiking these proteins at physiologically relevant concentrations into a rat TNF alpha positive serum. There was no crossreactivity detected.
• Standard Curve Range: 39.1 - 2,500 pg/mL
• Sensitivity: 11.0 pg/mL
• Assay Type: Sandwich ELISA
• Inter-Assay: CV% < 10%
• Intra-Assay: CV% < 5.0%
• Spike Recovery: 109%
• Sample Volume: 50 uL
• Kit Type: ELISA Kit (Colorimetric)
• Gene: TNF

Applications/Dilutions

• Dilutions:
  • ELISA

Packaging, Storage & Formulations

• Storage: Storage of components varies. See protocol for specific instructions.

Alternate Names for Rat TNF-alpha ELISA Kit (Colorimetric)

• APC1 protein
• Cachectin
• Cachetin
• DIF
• TNF
• TNF, monocyte-derived
• TNFA
• TNF-A
• TNFalpha
• TNF-alpha
• TNF-alphacachectin
• TNFATNF, macrophage-derived
• TNFG1F
• TNFSF1A
• TNFSF2
• TNFSF2TNF superfamily, member 2
• tumor necrosis factor (TNF superfamily, member 2)
• tumor necrosis factor alpha
• Tumor necrosis factor ligand superfamily member 2
• tumor necrosis factor
• tumor necrosis factor-alpha

Background

Tumor necrosis factor (TNF)-alpha is a pro-inflammatory cytokine belonging to the TNF superfamily that is secreted by monocytes/macrophages, T cell, and natural killer (NK), among others (1). TNF-alpha is synthesized as a 233 amino acid (aa) transmembrane protein (mTNF-alpha) with a theoretical molecular weight (MW) of 26 kDa (1,2) that forms a homotrimer. mTNF-alpha is cleaved by TNF-alpha converting enzyme (TACE) and released in its 157 aa, 17 kDa soluble form (sTNF-alpha) (1-5). Both mTNF-alpha and sTNF-alpha are capable of binding type 1 TNF receptors (TNFR1), whereas mTNF-alpha predominately binds to TNFR2 (1,2). TNF-alpha binding to its receptors causes receptor recruitment of adaptor proteins, formation of signaling complexes, and downstream signaling cascades (e.g. MAPK, NF-kappaB, and Caspase-8), leading to distinct cellular responses such as survival, proliferation, inflammation, necroptosis, and apoptosis (1-5).

TNF-alpha is critical for normal immune response; however, dysregulation of TNF-alpha production can result in various pathologies (2,4,5). Excessive production of pro-inflammatory cytokines including interleukin 1 (IL-1), IL-6, and TNF-alpha has been implicated in an array of autoimmune diseases like rheumatoid arthritis (RA), inflammatory bowel disease (IBD), and psoriasis (2,4,5). Anti-TNF monoclonal antibodies, including Infliximab, and soluble TNFR have been approved for the treatment of autoimmune and TNF-mediated diseases (5). Additionally, data suggests that TNF inhibitors can be beneficial for treating patients experiencing immune-related adverse events associated with immune checkpoint inhibitor cancer treatment (6).

References

1. Holbrook J, Lara-Reyna S, Jarosz-Griffiths H, McDermott M. Tumour necrosis factor signalling in health and disease. F1000Res. 2019;8:F1000 Faculty Rev-111. https://doi.org/10.12688/f1000research.17023.1

2. Jang DI, Lee AH, Shin HY, et al. The Role of Tumor Necrosis Factor Alpha (TNF-alpha) in Autoimmune Disease and Current TNF-alpha Inhibitors in Therapeutics. Int J Mol Sci. 2021;22(5):2719. https://doi.org/10.3390/ijms22052719

3. Horiuchi T, Mitoma H, Harashima S, Tsukamoto H, Shimoda T. Transmembrane TNF-alpha: structure, function and interaction with anti-TNF agents. Rheumatology (Oxford). 2010;49(7):1215-1228. https://doi.org/10.1093/rheumatology/keq031

4. Webster JD, Vucic D. The Balance of TNF Mediated Pathways Regulates Inflammatory Cell Death Signaling in Healthy and Diseased Tissues. Front Cell Dev Biol. 2020;8:365. https://doi.org/10.3389/fcell.2020.00365

5. Kalliolias GD, Ivashkiv LB. TNF biology, pathogenic mechanisms and emerging therapeutic strategies. Nat Rev Rheumatol. 2016; 12(1):49-62. https://doi.org/10.1038/nrrheum.2015.169

6. Chen AY, Wolchok JD, Bass AR. TNF in the era of immune checkpoint inhibitors: friend or foe?. Nat Rev Rheumatol. 2021;17(4):213-223. doi:10.1038/s41584-021-00584-4

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. ELISA Kits are guaranteed for 6 months from date of receipt.

Publications for TNF-alpha ELISA Kit (NBP1-92681)(10)

Publications using NBP1-92681

• Li L, Ma H, Zhang Y et al. Protocatechuic acid reverses myocardial infarction mediated by b-adrenergic agonist via regulation of Nrf2/HO-1 pathway, inflammatory, apoptotic, and fibrotic events Journal of biochemical and molecular toxicology 2023-01-02 [PMID: 36593721]
• Mohamed K, Abdelfattah M, El-khadragy M et al. Rutin-loaded selenium nanoparticles modulated the redox status, inflammatory, and apoptotic pathways associated with pentylenetetrazole-induced epilepsy in mice Green Processing and Synthesis 2023-01-26 (ELISA, Mouse)
• ALRashdi BM, Mohamed RA, Mohamed AH et al. Therapeutic activity of green synthesized selenium nanoparticles from turmeric against cisplatin-induced oxido-inflammatory stress and cell death in mice kidney Bioscience reports 2023-11-30 [PMID: 37902021] (ELISA)
• Li H, Yuan Y, Chen H et al. Indoleamine 2,3-dioxygenase mediates the therapeutic effects of adipose-derived stromal/stem cells in experimental periodontitis by modulating macrophages through the kynurenine-AhR-NRF2 pathway Molecular metabolism 2022-10-18 [PMID: 36270612] (ELISA)
• Alsharif K, Albrakati A, omairi N et al. The therapeutic antischizophrenic activity of prodigiosin and selenium co-supplementation against amphetamine hydrochloride-induced behavioural changes, oxidative, inflammatory, and apoptotic challenges in rats Research Square 2022-06-28 [PMID: 36048389] (ELISA)
• Al Omairi N, Albrakati A, Alsharif K et al. Selenium Nanoparticles with Prodigiosin Rescue Hippocampal Damage Associated with Epileptic Seizures Induced by Pentylenetetrazole in Rats Biology 2022-02-23 [PMID: 35336729]
• Albrakati A, Alsharif KF, Al Omairi NE et al. Neuroprotective Efficiency of Prodigiosins Conjugated with Selenium Nanoparticles in Rats Exposed to Chronic Unpredictable Mild Stress is Mediated Through Antioxidative, Anti-Inflammatory, Anti-Apoptotic, and Neuromodulatory Activities International journal of nanomedicine 2021-12-30 [PMID: 35002238]
• Liu Y, Shen W, Chen Q et al. Inhibition of RAGE by FPS-ZM1 alleviates renal injury in spontaneously hypertensive rats European Journal of Pharmacology 2020-06-01 [PMID: 32502492] (ELISA, Rat)
• Abdelfattah MS, Elmallah MIY, Ebrahim HY et al. Prodigiosins from a marine sponge-associated actinomycete attenuate HCl/ethanol-induced gastric lesion via antioxidant and anti-inflammatory mechanisms PLoS ONE 2019-06-13 [PMID: 31194753] (ELISA, Rat)
• Sun Y, Zhao DL, Liu ZX et al. Beneficial effect of 20-hydroxyecdysone exerted by modulating antioxidants and inflammatory cytokine levels in collagen-induced arthritis: A model for rheumatoid arthritis Mol Med Rep 2017-08-29 [PMID: 28901397] (ELISA, Rat)

FAQs for TNF-alpha ELISA Kit (NBP1-92681). (Showing 1 - 5 of 5 FAQs).

1. I need to know about TNF-alpha antibody kits for mice.
   • We currently have two TNFalpha ELISA kits specific for mouse: catalog numbers KA0257 and NBP1-92670.
2. I would like to ask you for help. I need an antibody for Elisa to detect human TNF alpha in a supernatant from a cell culture (meaning supernatant after centrifugation of collected cell suspension from a plate well). Which of your antibodies against human tnf alpha would be suitable? I would like to buy only the primary antibody.
   • I would recommend catalog number NBP1-67821 or NB600-587; however, a full list of our anti-human TNF alpha antibodies suitable for use in ELISA can be found using this link.
3. I am interested in a TNF alpha antibody, cross reactive for human, rat and mouse (host species: rabbit). Could your product NBP1-19532 be used in western blotting applications? Or do you have a similar product in your catalog which could fit with my request?
   • The antibody you mention, NBP1-19532, has not yet been validated in Western blot. I would instead recommend either NBP1-67821 or NB600-587. These are both rabbit polyclonal antibodies that cross-reacts with human, mouse and rat and have been used in Western blotting.
4. Can your TNF-alpha products be used to treat TBI victims and therefore avoid the perispinal injections with Enbrel?
   • I am very sorry, but all of our products are for scientific research use only, and none are intended or approved for use in humans.
5. Can we use NBP1-92681 (Rat TNF-alpha ELISA Kit, Biotin) on rat tissue lysate? If yes, please share how to prepare the lysate?
   • Yes, this kit is for rat samples especially the cell culture supernatant and serum but it should be good for rat tissue lysate also. We have not tested it on tissue lysates yet, but we have had reports from other customers using lysis buffers containing 0.5% TX-100 showing no deleterious effect on the performance of some of our kits. We do recommend that as a positive control you spike in some of the standard at a known concentration in your buffer and see if it performs as expected. In short, the lab recommended to use one or more cell line supernatants as the positive control in the ELISA assays.

Bioinformatics

• Gene Symbol: TNF