Mouse TNF-alpha ELISA

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


Antigen 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

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

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.


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. Trivedi PP., Jena GB., Tikoo KB., Kumar V. Melatonin Modulated Autophagy and Nrf2 Signaling Pathways in Mice With Colitis-Associated Colon Carcinogenesis. Molecular Carcinogenesis. Article first published online: 16 JAN 2015 | DOI: 10.1002/mc.22274
    Species: Mouse
    Sample type: Tissue Lysate
  2. Yu X., Yao JY., He J., Tian JW. Protection of MPTP-induced neuroinflammation and neurodegeneration by rotigotine-loaded microspheres. Life Sci. 2015 Mar 1;124:136-43. doi: 10.1016/j.lfs.2015.01.014
    Species: Mouse
    Sample type: Tissue Lysate
  3. Zhong E., Xu G., Wang Z., et al. Identification ofanti-inflammatoryconstituentsfrom Kalimerisindica with UHPLC-ESI-Q-TOF-MS/MSandGC-MS. J Ethnopharmacol. 2015 May 13;165:39-45. doi: 10.1016/j.jep.2015.02.034.
    Species: Mouse
    Sample type: Conditioned Media
  4. Ling Z., Liu X., Cheng Y., et al. Clostridium butyricum Combined with Bifidobacterium infantis Probiotic Mixture Restores Fecal Microbiota and Attenuates Systemic Inflammation in Mice with Antibiotic-Associated Diarrhea. Biomed Res Int. 2015; 2015: 582048. Published online 2015 Feb 23. doi: 10.1155/2015/582048
    Species: Mouse
    Sample type: Serum
  5. Singla D., Singla R., Abdelli L., Glass C. Fibroblast growth factor-9 enhances M2 macrophage differentiation and attenuates adverse cardiac remodeling in the infarcted diabetic heart. PLoS One. 2015 Mar 13;10(3):e0120739. doi: 10.1371/journal.pone.0120739
    Species: Mouse
    Sample type: Tissue Lysate
  6. Li J., Chai Q., Zhang Y., et al. Mycobacterium tuberculosis Mce3E suppresses host innate immune responses by targeting ERK1/2 signaling. J Immunol. 2015 Apr 15;194(8):3756-67. doi: 10.4049/jimmunol.1402679.
    Species: Mouse
    Sample type: Conditioned Media
  7. Jia Y., Zheng Z., Wang Y., et al. SIRT1 is a regulator in high glucose-induced inflammatory response in RAW264.7 cells. PLoS One. 2015 Mar 20;10(3):e0120849. doi: 10.1371/journal.pone.0120849
    Species: Mouse
    Sample type: Conditioned Media
  8. Galam L., Parthasarathy P., Cho Y., et al. Adenovirus-mediated transfer of the SOCS-1 gene to mouse lung confers protection against hyperoxic acute lung injury. Free Radic Biol Med. 2015 Jul;84:196-205. doi: 10.1016/j.freeradbiomed.2015.03.036.
    Species: Mouse
    Sample type: BAL Fluid
  9. Dey P., Dutta S., Sarkar MP., Chaudhuri T. Assessment of hepatoprotective potential of N. indicum leaf on haloalkane xenobiotic induced hepatic injury in Swiss albino mice. Chem Biol Interact. 2015 Jun 25;235:37-46. doi: 10.1016/j.cbi.2015.03.025.
    Species: Mouse
    Sample type: Conditioned Media
  10. Sun J., Wang F., Li H., et al. Neuroprotective Effect of Sodium Butyrate against Cerebral Ischemia/Reperfusion Injury in Mice. Biomed Res Int. 2015;2015:395895. doi: 10.1155/2015/395895.
    Species: Mouse
    Sample type: Tissue Lysate
I used the kit for cell culture supernatant analysis. The dilution times need yourself to investigate ahead and then do lots of experiments. Good products.

Mouse spleen lysates and with denervated muscle tissue was a sample for use of the kit. There's very little reaction with lysates but it exists. From my experience, this kit works well for cell culture supernatants.

The kit was packaged in an organized and clear manner, and it was easy to walk through each step of the procedure. I used the mouse IL-6, VEGF, and TNF-alpha kits for plasma as well as homogenized muscle samples. Each worked well, and I would be happy to use them again.

Houston Methodist Research Institute
This kit worked very well for the analysis of mouse serum samples. It was shipped quickly, easy to use, and did exactly what it was supposed to. I highly recommend this product!

BTS Research
I used this ELISA kit for Serum and cell culture supernatant analysis. Great product!
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This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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