Rat TNF-alpha ELISA

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

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

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Assay Format

Detection Range:
25 pg/ml - 20000 pg/ml
Recommended Dilution (serum & plasma):
2 fold
25 pg/ml

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This ELISA kit shows no cross-reactivity with any of the cytokines tested: rat CINC-2, CINC-3, CNTF, Fractalkine, IL-1 alpha, IL-1 beta, IL-4, IL-6, IL-10, GM-CSF, IFN-gamma, Leptin, Lix, MCP-1, MIP-3 alpha, beta- NGF, TIMP-1, VEGF.
Number of Targets Detected:
Species Detected:
  • Rat
Compatible Sample Types:
  • Cell Culture Supernatants
  • Cell Lysates
  • Plasma
  • Serum
  • Tissue Lysates
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:
  • P16599
Gene ID:
  • 24835
  • 103694380
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:
  • Rat

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.

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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. Oyewo, Emmanuel Bukoye, and Musbau Adewumi Akanji. "Immune modulation potentials of aqueous extract of Andrographis paniculata leaves in male rat."?Researcher?3.1 (2011): 48-57.
    Species: Rat
    Sample type: Serum
  2. Jourdan, Tony, et al. "Activation of the Nlrp3 inflammasome in infiltrating macrophages by endocannabinoids mediates beta cell loss in type 2 diabetes." Nature medicine 19.9 (2013): 1132.
    Species: Rat
    Sample type: Cell Lysate (diabetes model)
  3. Knapp, Darin J., et al. "Cytokine involvement in stress may depend on corticotrophin releasing factor to sensitize ethanol withdrawal anxiety." Brain, behavior, and immunity 25 (2011): S146-S154.
    Species: Rat
    Sample type: Tissue lysate
  4. Sachdeva C., Mishra N., Sharma S. Development and characterization of enteric-coated microparticles of biochanin A for their beneficial pharmacological potential in estrogen deficient-hypertension. Drug Deliv. 2016 Jul;23(6):2044-57. doi: 10.3109/10717544.2015.1114046.
    Species: Rat
    Sample type: Tissue Lysate (Aortic tissue lysate)
  5. Allabassi M. The Possible Protective Effects of Zileuton against Pulmonary Fibrosis Induced by Amiodarone in Male Rats. Journal of Natural Sciences Research. Vol.5, No.2, 2015
    Species: Rat
    Sample type: Tissue Lysate (Lung tissue treated with Zileuton)
  6. Abdelkader NF., Safar MM., Salem H. Ursodeoxycholic Acid Ameliorates Apoptotic Cascade in the Rotenone Model of Parkinson's Disease: Modulation of Mitochondrial Perturbations. Mol Neurobiol. 2014 Dec 15. [Epub ahead of print]. DOI 10.1007/s12035-014-9043-8
    Species: Rat
    Sample type: Tissue Lysate (Striatal brain tissue samples)
  7. Tang J., Chen Q., Guo J., et al. Minocycline Attenuates Neonatal Germinal-Matrix-Hemorrhage-Induced Neuroinflammation and Brain Edema by Activating Cannabinoid Receptor 2. Mol Neurobiol. 2015 Apr 2. epub ahead of print
    Species: Rat
    Sample type: Tissue Lysate (Brain lysates in matrix Hemorrhage-Induced mice)
  8. Hedya SA, Safar MM, Bahgat AK. Cilostazol Mediated Nurr1 and Autophagy Enhancement: Neuroprotective Activity in Rat Rotenone PD Model. Mol Neurobiol. 2018 Sep;55(9):7579-7587. doi: 10.1007/s12035-018-0923-1. Epub 2018 Feb 10. PMID: 29429051.
    Species: Rat
    Sample type: Tissue Lysate (Striatal Tissue)
  9. Ikutomi M., Sahara M., Nakajima T., et al. Diverse contribution of bone marrow-derived late-outgrowth endothelial progenitor cells to vascular repair under pulmonary arterial hypertension and arterial neointimal formation. J Mol Cell Cardiol. 2015 Sep;86:121-35. doi: 10.1016/j.yjmcc.2015.07.019.
    Species: Rat
    Sample type: Conditioned Media (Endothelial progenitor cells, Rat endothelial aoritc cells, AoECS, and BM-derived mononuclear cells)
  10. Chen B., Zhou W., Ning M., et al. Evaluation of antitumour activity of tea carbohydrate polymers in hepatocellular carcinoma animals. Int J Biol Macromol. 2012 May 1;50(4):1103-8. doi: 10.1016/j.ijbiomac.2012.03.001.
    Species: Human
    Sample type: Serum (Hepatocellular carcinoma rat model treated with tea carbohydrate)
We used the rat TNF-alpha ELISA kit to determine concentrations of TNF alpha in our rat plasma (heparin anticoagulated) and kidney extract samples. After determining optimal concentrations using two strips, ran the entire plate and everything worked perfectly. We were able to corroborated results from other experiments.

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This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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