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Bovine TNF-alpha ELISA

RayBio® Bovine 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:
0.1 ng/ml - 30 ng/ml
Recommended Dilution (serum & plasma):
2 fold
0.1 ng/ml

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This ELISA antibody pair detects bovine TNF-alpha. Other species not determined
Number of Targets Detected:
Species Detected:
  • Bovine (cow)
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:
  • Q06599
Gene ID:
  • 280943
Gene Symbols:
  • TNF
  • TNFA
  • 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:
  • Bovine (cow)

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. Zhang L., Huang C., Guo Y., et al. MicroRNA-26b Modulates the NF-?B Pathway in Alveolar Macrophages by Regulating PTEN. J Immunol. 2015 Dec 1;195(11):5404-14. doi: 10.4049/jimmunol.1402933.
    Species: Bovine
    Sample type: Conditioned Media (Bovine alveolar macrophages treated with LPS of M. haemolytica LPS)
  2. Li J., Hu L., Liu Y., et al. DDX19A Senses Viral RNA and Mediates NLRP3-Dependent Inflammasome Activation. J Immunol. 2015 Dec 15;195(12):5732-49. doi: 10.4049/jimmunol.1501606
    Species: Porcine
    Sample type: Conditioned Media (Primary pulmonary alveolar macrophages infected with porcine reproductive respt virus)
  3. Brandao A., Cooke R., Corra F., et al. Physiologic, health, and production responses of dairy cows supplemented with an immunomodulatory feed ingredient during the transition period. J Dairy Sci. 2016 Jul;99(7):5562-72. doi: 10.3168/jds.2015-10621
    Species: Bovine
    Sample type: Serum (LPS challenged cows)
  4. Rodrigues M., Cooke R., Marques R., et al. Effects of vaccination against respiratory pathogens on feed intake, metabolic, and inflammatory responses in beef heifers. J Anim Sci. 2015 Sep;93(9):4443-52. doi: 10.2527/jas.2015-9277.
    Species: Bovine
    Sample type: Serum (respiratory vaccine studies in beef heifers)
  5. Rodrigues M., Cooke R., Marques R., et al. Effects of oral meloxicam administration to beef cattle receiving lipopolysaccharide administration or vaccination against respiratory pathogens. J Anim Sci. 2015 Oct;93(10):5018-27. doi: 10.2527/jas.2015-9424.
    Species: Bovine
    Sample type: Serum (LPS and vaccinated cattle testing)

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

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