Mouse TNF-alpha ELISA

RayBio® Mouse TNF alpha ELISA Kit for cell culture supernatants, plasma, and serum samples.

Lead time: Typically ships within 1-2 business days. No Friday shipments.


Assay Format

Detection Range:
60 pg/ml - 6000 pg/ml
Recommended Dilution (serum & plasma):
2 fold
60 pg/ml

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This ELISA kit shows no cross-reactivity with any of the cytokines tested: Mouse CD30, L CD30, T CD40, CRG-2, CTACK, CXCL16, Eotaxin , Eotaxin-2, Fas Ligand, Fractalkine, GCSF, GM-CFS, IFN- gamma, IGFBP-3, IGFBP-5, IGFBP-6, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-3 Rb, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 p40/p70, IL-12 p70, IL-13, IL-17, KC, Leptin R, LEPTIN(OB), LIX, L-Selectin, Lymphotactin, MCP-1, MCP-5, M-CSF, MIG, MIP-1 alpha, MIP-1 gamma, MIP-2, MIP-3 beta, MIP-3 alpha, PF-4, P-Selectin, RANTES, SCF, SDF-1 alpha, TARC, TCA-3, TECK, TIMP-1, TNF RI, TNF RII, TPO, VCAM-1, VEGF.
Number of Targets Detected:
Species Detected:
  • Mouse
Compatible Sample Types:
  • Cell Culture Supernatants
  • Plasma
  • Serum
Design Principle:
  • Sandwich-based
Method of Detection:
  • Quantitative
Solid Support:
96-well Microplate

Product Specifications

1, 2, or 5 x 96-Well Strip Microplate Kit

Protein Information

Accession Number:
  • P06804
Gene ID:
  • 21926
Gene Symbols:
  • TNF
  • TNFSF2
Protein Name & Synonyms:
Tumor necrosis factor (Cachectin) (TNF-alpha) (Tumor necrosis factor ligand superfamily member 2) (TNF-a) [Cleaved into: Tumor necrosis factor, membrane form (N-terminal fragment) (NTF) Intracellular domain 1 (ICD1) Intracellular domain 2 (ICD2) C-domain 1 C-domain 2 Tumor necrosis factor, soluble form]
Target Species:
  • Mouse

Target Background

Tumor necrosis factor alpha (TNF alpha), also known as lymphotoxin and cachectin, is a pleiotropic cytokine. TNF alpha is a smaller cytokine with a molecular weight of 26 kDa (transmembrane) and 17 kDa (soluble) that binds to the same receptors producing a vast array of effects similar to those of TNF-beta. Both TNF-alpha and TNF-beta share 30% amino acid homology and have similar biological activities. TNF-alpha is produced primarily by monocytes, neutrophils and macrophages. It is found in synovial cells and macrophages in the tissues. TNF-alpha and TNF-beta can modulate many immune and inflammatory functions, while having the ability to inhibit tumor growth. Target tumor cells must express TNF receptors 1 (55 kDa) and 2 (75 kDa) to be killed, with the p55 receptor mediating the cytotoxic response.

Product Features

Additional Notes
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data

Standard Curves

Spiking & Recovery Results

Linearity Results

Application Notes

Kit Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Other Materials Required
  • Distilled or deionized water
  • Precision pipettes to deliver 2 µl to 1 µl volumes
  • Adjustable 1-25 µl pipettes for reagent preparation
  • 100 µl and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis
Protocol Outline
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 µl of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4°C.
  4. Add 100 µl of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.

Need your results faster? Try RayBiotech's SpeedELISA platform for quantitative detection in just three hours.


The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
  1. Fentahun Darge, H., Yibru Hanurry, E., Simegniew Birhan, Y., Worku Mekonnen, T., Tizazu Andrgie, A., Chou, H.-Y., … Tsai, H.-C. (2020). Multifunctional drug-loaded micelles encapsulated in thermo-sensitive hydrogel for in vivo local cancer treatment: synergistic effects of anti-vascular and immuno-chemotherapy. Chemical Engineering Journal, 126879. doi:10.1016/j.cej.2020.126879 
    Species: Mouse
    Sample type: Serum
  2. Škandík, M., Mrvová, N., Bezek, Š., & Ra?ková, L. (2020). Semisynthetic quercetin-quinone mitigates BV-2 microglia activation through modulation of Nrf2 pathway. Free Radical Biology and Medicine. doi:10.1016/j.freeradbiomed.2020.02.030 
    Species: Mouse
    Sample type: Conditioned media
  3. Liu N , Zhang GX , Niu YT , Wang Q , Zheng J , Yang JM , Sun T , Niu JG , Yu JQ . Anti-inflammatory and analgesic activities of indigo through regulating the IKK?/I?B/NF-?B pathway in mice. Food Funct. 2020 Oct 1;11(10):8537-8546. doi: 10.1039/c9fo02574j. Epub 2020 Oct 14. PMID: 33084638.
    Species: Mouse
    Sample type: Tissue Lysate
  4. Jia H, Guo J, Wang P, Sun K, Chen J, Ren W, Wei T, Yang Y, Li J, Liu X, Li R, Zhong J, Wang M, Tian Z, Feng Z, Zhao T. A self-designed CpG ODN enhanced the anti-melanoma effect of pimozide. Int Immunopharmacol. 2020 Jun;83:106397. doi: 10.1016/j.intimp.2020.106397. Epub 2020 Mar 25. PMID: 32220805.
    Species: Mouse
    Sample type: Conditioned Media
  5. Drolia R, Amalaradjou MAR, Ryan V, Tenguria S, Liu D, Bai X, Xu L, Singh AK, Cox AD, Bernal-Crespo V, Schaber JA, Applegate BM, Vemulapalli R, Bhunia AK. Receptor-targeted engineered probiotics mitigate lethal Listeria infection. Nat Commun. 2020 Dec 11;11(1):6344. doi: 10.1038/s41467-020-20200-5. PMID: 33311493; PMCID: PMC7732855.
    Species: Mouse
    Sample type: Tissue Lysate
  6. Gu I, Brownmiller C, Stebbins NB, Mauromoustakos A, Howard L, Lee SO. Berry Phenolic and Volatile Extracts Inhibit Pro-Inflammatory Cytokine Secretion in LPS-Stimulated RAW264.7 Cells through Suppression of NF-?B Signaling Pathway. Antioxidants (Basel). 2020 Sep 15;9(9):871. doi: 10.3390/antiox9090871. PMID: 32942640; PMCID: PMC7554842.
    Species: Mouse
    Sample type: Conditioned Media
  7. Scarcello E, Abdel-Mottaleb MMA, Beduneau A, Moulari B, Pellequer Y. Amelioration of murine experimental colitis using biocompatible cyclosporine A lipid carriers. Drug Deliv Transl Res. 2020 Aug 13. doi: 10.1007/s13346-020-00835-z. Epub ahead of print. PMID: 32794147.
    Species: Mouse
    Sample type: Conditioned Media
  8. Sharma P, Kulkarni GT, Sharma B. Possible involvement of D2/D3 receptor activation in ischemic preconditioning mediated protection of the brain. Brain Res. 2020 Dec 1;1748:147116. doi: 10.1016/j.brainres.2020.147116. Epub 2020 Sep 11. PMID: 32919985.
    Species: Mouse
    Sample type: Conditioned Media
  9. Xia H, Ge Y, Wang F, Ming Y, Wu Z, Wang J, Sun S, Huang S, Chen M, Xiao W, Yao S. Protectin DX ameliorates inflammation in sepsis-induced acute lung injury through mediating PPAR?/NF-?B pathway. Immunol Res. 2020 Oct;68(5):280-288. doi: 10.1007/s12026-020-09151-7. Epub 2020 Aug 26. PMID: 32845434.
    Species: Mouse
    Sample type: BAL Fluid
  10. Wang J, Wang B, Lv X, Wang L. NIK inhibitor impairs chronic periodontitis via suppressing non-canonical NF-?B and osteoclastogenesis. Pathog Dis. 2020 Oct 7;78(7):ftaa045. doi: 10.1093/femspd/ftaa045. PMID: 32860691.
    Species: Mouse
    Sample type: Conditioned Media

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