Rat TNF-alpha ELISA

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

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Antigen 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

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

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. Patyar, Sazal, et al. "Chemopreventive effect of artesunate in 1, 2-dimethylhydrazine-induced rat colon carcinogenesis."?Journal of advanced pharmaceutical technology & research8.3 (2017): 102.
    Species: Rat
    Sample type: plasma
  2. Dokubo, A., A. A. Uwakwe, and B. A. Amadi. "A study on the antioxidant and anti-inflammatory effects of aframomum sceptrum and parinari congensis seed extracts in alloxan-induced diabetic wistar rats."?EPRA International Journal of Research & Development?2.7 (2017): 1-9.
    Species: Rat
    Sample type: Tissue
  3. Xu B., Yang H., Sun M., et al. 2,3',4,4',5-Pentachlorobiphenyl Induces Inflammatory Responses in the Thyroid Through JNK and Aryl Hydrocarbon Receptor-Mediated Pathway. Toxicol Sci. 2016 Feb;149(2):300-11. doi: 10.1093/toxsci/kfv235.
    Species: Rat
    Sample type: Conditioned Media
  4. Rana A., Sharma S. Mechanism of sphingosine-1-phosphate induced cardioprotection against I/R injury in diabetic rat heart: Possible involvement of glycogen synthase kinase 3? and mitochondrial permeability transition pore. Clin Exp Pharmacol Physiol. 2016 Feb;43(2):166-73. doi: 10.1111/1440-1681.12516.
    Species: Rat
    Sample type: Tissue Lysate
  5. 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
  6. El-Mahdy N., Salem M., El-Sayad M., et al. Bone marrow mononuclear cells enhance anti-inflammatory effects of pravastatin against isoproterenol-induced myocardial infarction in rats. J Immunotoxicol. 2016 May;13(3):393-402. doi: 10.3109/1547691X.2015.1111959.
    Species: Rat
    Sample type: Tissue lysate
  7. Ram M., Singh V., Kumawat S., et al. Bilirubin modulated cytokines, growth factors and angiogenesis to improve cutaneous wound healing process in diabetic rats. Int Immunopharmacol. 2016 Jan;30:137-49. doi: 10.1016/j.intimp.2015.11.037
    Species: Rat
    Sample type: Tissue lysate
  8. Wang Y., Chen J., Luo X., et al. Ginsenoside metabolite compound K exerts joint-protective effect by interfering with synoviocyte function mediated by TNF-? and Tumor necrosis factor receptor type 2. Eur J Pharmacol. 2016 Jan 15;771:48-55. doi: 10.1016/j.ejphar.2015.12.019.
    Species: Rat
    Sample type: Conditioned Media
  9. Li P., Liu H., Sun P., et al. Chronic vagus nerve stimulation attenuates vascular endothelial impairments and reduces the inflammatory profile via inhibition of the NF-?B signaling pathway in ovariectomized rats. Exp Gerontol. 2016 Feb;74:43-55. doi: 10.1016/j.exger.2015.12.005.
    Species: Rat
    Sample type: Serum
  10. Abdel kawy H. Cilostazol attenuatescholestaticliverinjuryanditscomplications in commonbileductligatedrats. Eur J Pharmacol. 2015 Apr 5;752:8-17. doi: 10.1016/j.ejphar.2015.01.044.
    Species: Rat
    Sample type: Tissue Lysate
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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